Methods for diagnosing and evaluating cancer

ABSTRACT

Methods for diagnosing cancer, such as prostate, ovarian or breast cancer, as well as leukemia, are provided. Such methods may employ binding agents, such as antibodies or CAR sequences of OB-cadherin or N-cadherin, that specifically bind to OB-cadherin or N-cadherin, or polynucleotides that hybridize to a polynucleotide encoding OB-cadherin or N-cadherin. Also provided are methods for monitoring the progression of a cancer and to evaluate the metastatic potential of a cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. Ser. No. 09/234,395,filed Jan. 20, 1999; which is a continuation-in-part of U.S. Ser. No.09/187,859, filed Nov. 6, 1998; which is a continuation-in-part of U.S.Ser. No. 09/073,040, filed May 5, 1998.

TECHNICAL FIELD

[0002] The present invention relates generally to methods for cancerdiagnosis, and more particularly to the use of compounds that detectexpression of OB-cadherin or N-cadherin for diagnosing and determiningthe metastatic potential of cancers such as breast, ovarian and prostatecancer, as well as leukemia.

BACKGROUND OF THE INVENTION

[0003] Cancer is a significant health problem throughout the world.Although advances have been made in detection and therapy of cancer, novaccine or other universally successful method for prevention ortreatment is currently available. For example, among women, breast andovarian cancer are prevalent in the United States and other countries.Breast cancer, in particular, remains the second leading cause ofcancer-related deaths in women, affecting more than 180,000 women in theUnited States each year. For women in North America, the life-time oddsof getting breast cancer are now one in eight. Management of the diseasecurrently relies on a combination of early diagnosis (through routinebreast screening procedures) and aggressive treatment, which may includeone or more of a variety of treatments such as surgery, radiotherapy,chemotherapy and hormone therapy. The course of treatment for aparticular breast cancer is often selected based on a variety ofprognostic parameters, including an analysis of specific tumor markers.See, e.g., Porter-Jordan and Lippman, Breast Cancer 8:73-100, 1994.However, it remains difficult to evaluate the metastatic potential of acancer, and the high mortality observed in breast cancer patientsindicates that improvements are needed in the diagnosis and managementof the disease.

[0004] Prostate cancer is the most common form of cancer among males,with an estimated incidence of 30% in men over the age of 50. Humanprostate cancer has the propensity to metastasize to bone. Treatment iscommonly based on surgery and/or radiation therapy, but these methodsare ineffective in a significant percentage of cases, and this prevalentdisease is currently the second leading cause of cancer death among menin the U.S. To improve treatment of the disease, early diagnosis iscritical, but prostate cancer remains difficult to detect accurately.Two prostate specific proteins, prostate specific antigen (PSA) andprostatic acid phosphatase (PAP), have been used for diagnosis, buttechniques employing such proteins cannot provide complete diagnosticinformation. For example, PSA measurements not indicate the level ofmetastasis of a prostate cancer.

[0005] Although additional markers for prostate and other cancerscontinue to be discovered, there is presently no accurate method forevaluating the metastatic potential of these cancers. In order toimprove cancer treatment and survival, techniques that permit a moreaccurate diagnosis are needed. The present invention fulfills theseneeds and further provides other related advantages.

SUMMARY OF THE INVENTION

[0006] Briefly stated, this invention provides compositions and methodsfor diagnosing cancer, such as breast, ovarian and prostate cancer, aswell as leukemia. Certain methods provided herein employ binding agents,such as antibodies and fragments thereof, that specifically recognizeOB-cadherin or N-cadherin. Other methods employ one or morepolynucleotides capable of hybridizing to a polynucleotide encodingOB-cadherin or N-cadherin.

[0007] Within certain aspects, the present invention provides methodsfor determining the presence or absence of a cancer in a patient,comprising the steps of: (a) contacting a biological sample obtainedfrom a patient with a binding agent that specifically binds toOB-cadherin or N-cadherin; and (b) detecting in the sample an amount ofpolypeptide that binds to the binding agent, relative to a predeterminedcut-off value, and therefrom determining the presence or absence of acancer in the patient.

[0008] Within further aspects, methods are provided for monitoring theprogression of a cancer in a patient, comprising the steps of: (a)contacting a biological sample obtained from a patient at a first pointin time with a binding agent that specifically binds to OB-cadherin orN-cadherin; (b) detecting in the sample an amount of polypeptide thatbinds to the binding agent; (c) repeating steps (a) and (b) using abiological sample obtained from the patient at a subsequent point intime; and (d) comparing the amount of polypeptide detected in step (c)to the amount detected in step (b) and therefrom monitoring theprogression of the cancer in the patient.

[0009] Within other aspects, methods are provided for evaluating themetastatic potential of a cancer in a patient, comprising the steps of:(a) contacting a biological sample obtained from a patient afflictedwith cancer with a binding agent that specifically binds to OB-cadherinor N-cadherin; and (b) detecting in the sample an amount of polypeptidethat binds to the binding agent, relative to a predetermined cut-offvalue, and therefrom evaluating the metastatic potential of the cancerin the patient.

[0010] Kits for determining the presence or absence of a cancer in apatient are also provided. Such kits may comprise: (a) an antibody orantigen-binding fragment thereof that specifically binds to anOB-cadherin or N-cadherin CAR sequence; and (b) a detection reagent.

[0011] The present invention further provides methods for determiningthe presence or absence of a metastatic cancer in a patient, comprisingthe steps of: (a) contacting a biological sample obtained from a patientwith an oligonucleotide that hybridizes to a polynucleotide encodingOB-cadherin or N-cadherin; and (b) detecting in the sample a level of apolynucleotide that hybridizes to the oligonucleotide, relative to apredetermined cut-off value, and therefrom determining the presence orabsence of a metastatic cancer in the patient. Within certainembodiments, the amount of mRNA is detected via polymerase chainreaction using, for example, at least one oligonucleotide primer thathybridizes to a polynucleotide that encodes OB-cadherin or N-cadherin,or a complement of such a polynucleotide. Within other embodiments, theamount of mRNA is detected using a hybridization technique, employing anoligonucleotide probe that hybridizes to a polynucleotide that encodesOB-cadherin or N-cadherin, or a complement of such a polynucleotide. Ina preferred embodiment, at least one of the oligonucleotide primerscomprises at least about 10 contiguous nucleotides of a DNA moleculeencoding OB-cadherin or N-cadherin.

[0012] In related aspects, methods are provided for monitoringprogression of a cancer in a patient, comprising the steps of: (a)contacting a biological sample obtained from a patient with anoligonucleotide that hybridizes to a polynucleotide encoding OB-cadherinor N-cadherin; (b) detecting in the sample an amount of polynucleotidethat hybridizes to the oligonucleotide; (c) repeating steps (a) and (b)using a biological sample obtained from the patient at a subsequentpoint in time; and (d) comparing the amount of polynucleotide detectedin step (c) with the amount detected in step (b) and therefrommonitoring progression of a cancer in the patient.

[0013] Within other aspects, methods are provided for evaluating themetastatic potential of a cancer in a patient, comprising the steps of:(a) contacting a biological sample obtained from a patient with anoligonucleotide that hybridizes to a polynucleotide encoding OB-cadherinor N-cadherin; and (b) detecting in the sample an amount of apolynucleotide that hybridizes to the oligonucleotide, relative to apredetermined cut-off value, and therefrom evaluating the metastaticpotential of the cancer in the patient.

[0014] In related aspects, diagnostic kits comprising the aboveoligonucleotide probes or primers are provided.

[0015] These and other aspects of the present invention will becomeapparent upon reference to the following detailed description andattached drawings. All references disclosed herein are herebyincorporated by reference in their entirety as if each was incorporatedindividually.

BRIEF DESCRIPTION OF THE DRAWINGS

[0016]FIG. 1A is a diagram depicting the structure of classical CADs.The five extracellular domains are designated EC1-EC5, the hydrophobicdomain that transverses the plasma membrane (PM) is represented by TM,and the two cytoplasmic domains are represented by CP1 and CP2. Thecalcium binding motifs are shown by DXNDN (SEQ ID NO:1), DXD and LDRE(SEQ ID NO:2). The CAR sequence, HAV, is shown within EC1. Cytoplasmicproteins β-catenin (β), α-catenin (α) and α-actinin (ACT), which mediatethe interaction between CADs and microfilaments (MF) are also shown.FIG. 1B is a diagram depicting the structure of the atypical CAD knownas OB-cadherin. The CAR sequence, IFVIDDKSG (SEQ ID NO:3), is shownwithin EC 1.

[0017]FIG. 2 provides the amino acid sequences of representativemammalian OB-cadherin EC1 domains: human OB-cadherin (SEQ ID NO:4) andmouse OB-cadherin (SEQ ID NO:5).

[0018]FIG. 3 provides the amino acid sequences of mammalian classicalcadherin EC1 domains: human N-cadherin (SEQ ID NO:6), mouse N-cadherin(SEQ ID NO:7) and cow N-cadherin (SEQ ID NO:8).

[0019]FIGS. 4A-4C are photographs showing cultures of human breastcancer cells in the presence (FIGS. 4B and 4C) and absence (FIG. 4A) ofa representative linear OB-cadherin peptide. FIG. 4A shows the cells 24hours after exposure to 100 μl water/1 ml culture medium (magnification200×). FIGS. 4B and 4C show the cells 24 hours after exposure to 100 μLof a solution containing 10 mg/mL N-Ac-IFVIDDKSG-NH₂ (SEQ ID NO:9) per 1mL culture medium (magnifications of 200× and 100×, respectively).Arrows indicate rounded cells.

[0020]FIG. 5 is a photograph illustrating the results of PCR analysis todetect the presence of OB-cadherin in metastatic human ovarian cancercells, but not in well-differentiated human ovarian cancer cells. RT-PCRproducts from two cell lines are shown: SKOV3 in lane 1 and OVCAR3 inlane 2. The primers used were specific for OB-cadherin (OB-cad) andhypoxanthine phosphoribosyltransferase (HPRT) as indicated, with anexpected PCR product of 745 bp and 352 bp, respectively. Products werestained with ethidium bromide and resolved by agarose gelelectrophoresis. Lane M represents a 1 kb ladder (Gibco/BRL).

[0021]FIG. 6 is a photograph illustrating the results of PCR analysisdetecting the presence of OB-cadherin in leukemic cells. RT-PCR productswere generated from lymphocytes of a human B-CLL patient (lane I) andmouse liver (lane 2). The primers used were specific for OB-cadherin(OB-cad, top panel) and hypoxanthine phosphoribosyltransferase (HPRT,bottom panel), with an expected PCR product of 745 bp and 352 bp,respectively. Products were stained with ethidium bromide and resolvedby agarose gel electrophoresis. Lane M represents a 1 kb ladder(Gibco/BRL).

DETAILED DESCRIPTION OF THE INVENTION

[0022] As noted above, the present invention provides methods fordetecting and evaluating cancer in a patient. The methods providedherein are based, in part, on the discovery that OB-cadherin andN-cadherin are expressed by metastatic carcinoma cells, but not byhighly differentiated, poorly invasive carcinomas. Accordingly,metastatic cancers may be detected and monitored using methods thatevaluate OB-cadherin and/or N-cadherin expression. Certain methodsprovided herein employ binding agents, such as antibodies and fragmentsthereof, that specifically recognize an OB-cadherin or N-cadherinextracellular domain, or a portion thereof. Other methods employ one ormore polynucleotides that hybridize to a polynucleotide encodingOB-cadherin or N-cadherin, or to a sequence that is a complement of sucha polynucleotide. In general, within the methods provided herein, abiological sample obtained from a patient is typically contacted withsuch a binding agent or polynucleotide so as to permit determination ofthe level of OB-cadherin or N-cadherin, or the level of polynucleotideencoding OB-cadherin or N-cadherin within the sample. This determinationis indicative of the presence or absence of a metastatic cancer, and maybe used to evaluate the metastatic potential of a cancer and monitorcancer progression.

[0023] OB-Cadherin and N-Cadherin

[0024] As used herein, the terms “OB-cadherin” and “N-cadherin” refer tocell adhesion molecules that are expressed by a human or non-humanindividual, and that are substantially homologous to a known OB-cadherinor N-cadherin (OB-cadherin and N-cadherin are discussed, for example, inMunro et al., In: Cell Adhesion and Invasion in Cancer Metastasis, P.Brodt, ed., pp. 17-34, RG Landes Co., Austin Tex., 1996; OB-cadherin isalso described by Getsios et al., Developmental Dynamics 211:238-247,1998; Simonneau et al., Cell Adhesion and Communication 3:115-130, 1995;Okazaki et al., J. Biological Chemistry 269:12092-12098, 1994). CertainOB-cadherin molecules comprise a sequence provided in FIG. 2 and certainN-cadherin molecules comprise a sequence provided in FIG. 3, but thepresent invention also contemplates the use of OB-cadherin andN-cadherin sequences from other organisms. Such OB-cadherin andN-cadherin sequences may generally be identified based upon sequencesimilarity to the sequences provided herein and based upon the presenceof OB-cadherin or N-cadherin activity, using an assay provided herein.

[0025] For generating and testing binding agents, as described herein,polypeptides comprising a portion of OB-cadherin or N-cadherin may beused. Such polypeptides may, if they comprise a cell adhesionrecognition sequence (CAR sequence), also be used as binding agentsthemselves. Preferred portions are extracellular domains and portionsthereof, such as a portion comprising a CAR sequence. An extracellulardomain of an OB-cadherin or N-cadherin may generally be identified basedon homology to the extracellular domain sequences provided herein, andusing well known techniques, such as the presence of one or more of: ahydrophilic sequence, a region that is recognized by an antibody, aregion that is cleaved by trypsin and/or a potential glycosylation sitewith the glycosylation motif Asn-X-Ser/Thr. A polypeptide may comprisean entire extracellular domain or a portion thereof, and may consistentirely of the OB-cadherin or N-cadherin sequence, or may additionallycomprise further peptide and/or non-peptide regions. Additional peptideregions may be derived from OB-cadherin or N-cadherin and/or may beheterologous. Such a peptide may be a linear or cyclic peptide, cyclizedvia an intramolecular bond that may be backbone to backbone, side-chainto backbone or side-chain to side-chain.

[0026] Polypeptides comprising an OB-cadherin or N-cadherin sequence maybe synthesized by methods well known in the art, including chemicalsynthesis and recombinant DNA methods. For peptides up to about 50residues in length, chemical synthesis may be performed using solutionor solid phase peptide synthesis techniques, in which a peptide linkageoccurs through the direct condensation of the α-amino group of one aminoacid with the α-carboxy group of the other amino acid with theelimination of a water molecule. Peptide bond synthesis by directcondensation, as formulated above, requires suppression of the reactivecharacter of the amino group of the first amino acid and of the carboxylgroup of the second amino acid. The masking substituents must permittheir ready removal, without inducing breakdown of the labile peptidemolecule.

[0027] In solution phase synthesis, a wide variety of coupling methodsand protecting groups may be used (see Gross and Meienhofer, eds., “ThePeptides: Analysis, Synthesis, Biology,” Vol. 1-4 (Academic Press,1979); Bodansky and Bodansky, “The Practice of Peptide Synthesis,” 2ded. (Springer Verlag, 1994)). In addition, intermediate purification andlinear scale up are possible. Those of ordinary skill in the art willappreciate that solution synthesis requires consideration of main chainand side chain protecting groups and activation method. In addition,careful segment selection is necessary to minimize racemization duringsegment condensation. Solubility considerations are also a factor.

[0028] Solid phase pep tide synthesis uses an insoluble polymer forsupport during organic synthesis. The polymer-supported peptide chainpermits the use of simple washing and filtration steps instead oflaborious purifications at intermediate steps. Solid-phase peptidesynthesis may generally be performed according to the method ofMerrifield et al., J. Am. Chem. Soc. 85:2149, 1963, which involvesassembling a linear peptide chain on a resin support using protectedamino acids. Solid phase peptide synthesis typically utilizes either theBoc or Fmoc strategy. The Boc strategy uses a 1% cross-linkedpolystyrene resin. The standard protecting group for α-amino functionsis the tert-butyloxycarbonyl (Boc) group. This group can be removed withdilute solutions of strong acids such as 25% trifluoroacetic acid (TFA).The next Boc-amino acid is typically coupled to the amino acyl resinusing dicyclohexylcarbodiimide (DCC). Following completion of theassembly, the peptide-resin is treated with anhydrous HF to cleave thebenzyl ester link and liberate the free peptide. Side-chain functionalgroups are usually blocked during synthesis by benzyl-derived blockinggroups, which are also cleaved by HF. The free peptide is then extractedfrom the resin with a suitable solvent, purified and characterized.Newly synthesized peptides can be purified, for example, by gelfiltration, HPLC, partition chromatography and/or ion-exchangechromatography, and may be characterized by, for example, massspectrometry or amino acid sequence analysis. In the Boc strategy,C-terminal amidated peptides can be obtained using benzhydrylamine ormethylbenzhydrylamine resins, which yield peptide amides directly uponcleavage with HF.

[0029] In the procedures discussed above, the selectivity of theside-chain blocking groups and of the peptide-resin link depends uponthe differences in the rate of acidolytic cleavage. Orthoganol systemshave been introduced in which the side-chain blocking groups and thepeptide-resin link are completely stable to the reagent used to removethe α-protecting group at each step of the synthesis. The most common ofthese methods involves the 9-fluorenylmethyloxycarbonyl (Fmoc) approach.Within this method, the side-chain protecting groups and thepeptide-resin link are completely stable to the secondary amines usedfor cleaving the N-α-Fmoc group. The side-chain protection and thepeptide-resin link are cleaved by mild acidolysis. The repeated contactwith base makes the Merrifield resin unsuitable for Fmoc chemistry, andp-alkoxybenzyl esters linked to the resin are generally used.Deprotection and cleavage are generally accomplished using TFA.

[0030] Those of ordinary skill in the art will recognize that, in solidphase synthesis, deprotection and coupling reactions must go tocompletion and the side-chain blocking groups must be stable throughoutthe entire synthesis. In addition, solid phase synthesis is generallymost suitable when peptides are to be made on a small scale.

[0031] If desired, acetylation of the N-terminus can be accomplished byreacting the final peptide with acetic anhydride before cleavage fromthe resin. C-amidation is accomplished using an appropriate resin suchas methylbenzhydrylamine resin using the Boc technology. Followingsynthesis of a linear peptide, with or without N-acetylation and/orC-amidation, cyclization may be achieved, if desired, by any of avariety of techniques well known in the art.

[0032] For longer polypeptides, recombinant methods are preferred forsynthesis. Within such methods, all or part of a polypeptide can besynthesized in living cells, using any of a variety of expressionvectors known to those of ordinary skill in the art to be appropriatefor the particular host cell. Suitable host cells may include bacteria,yeast cells, mammalian cells, insect cells, plant cells, algae and otheranimal cells (e.g., hybridoma, CHO, myeloma). The DNA sequencesexpressed in this manner may be prepared based on known cDNA or genomicsequences, or from sequences isolated by screening an appropriatelibrary with probes designed based on the sequences of knownnonclassical cadherins. Such screens may generally be performed asdescribed in Sambrook et al., Molecular Cloning: A Laboratory Manual,Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y., 1989 (andreferences cited therein). Polymerase chain reaction (PCR) may also beemployed, using oligonucleotide primers in methods well known in theart, to isolate nucleic acid molecules encoding all or a portion of anendogenous adhesion molecule. To generate a nucleic acid moleculeencoding a desired polypeptide, an endogenous OB-cadherin or N-cadherinsequence may be modified using well known techniques. Alternatively,portions of the desired nucleic acid sequences may be synthesized usingwell known techniques, and then ligated together to form a sequenceencoding the polypeptide.

[0033] Proteins and peptides may be evaluated for OB-cadherin orN-cadherin activity using any of a variety of assays. An initial screenfor OB-cadherin or N-cadherin activity may be performed by evaluatingthe ability to bind to OB-cadherin or N-cadherin using any binding assayknown to those of ordinary skill in the art. For example, a PharmaciaBiosensor machine may be used, as discussed in Jonsson et al.,Biotechniques 11:520-27, 1991. A specific example of a technology thatmeasures the interaction of peptides with molecules can be found inWilliams et al., J. Biol. Chem. 272, 22349-22354, 1997. Alternatively,real-time BIA (Biomolecular Interaction Analysis) uses the opticalphenomenon surface plasmon resonance to monitor biomolecularinteractions. The detection depends upon changes in the massconcentration of macromolecules at the biospecific interface, which inturn depends upon the immobilization of test molecule or peptide(referred to as the ligand) to the surface of a Biosensor chip, followedby binding of the interacting molecule (referred to as the analyte) tothe ligand. Binding to the chip is measured in real-time in arbitraryunits of resonance (RU).

[0034] By way of example, surface plasmon resonance experiments may becarried out using a BIAcore X™ Biosensor (Pharmacia Ltd., BIAcore,Uppsala, Sweden). Parallel flow cells of CM 5 sensor chips may bederivatized, using the amine coupling method, with streptavidin (200μg/ml) in 10 mM Sodium Acetate, pH 4.0, according to the manufacturer'sprotocol. Approximately 2100-2600 resonance units (RU) of ligand may beimmobilized, corresponding to a concentration of about 2.1-2.6 ng/mm².The chips may then coated be with OB-cadherin or N-cadherin derivatizedto biotin. Any non-specifically bound protein is removed.

[0035] To determine binding, test analytes (e.g., peptides or proteins)may be placed in running buffer and passed simultaneously over test andcontrol flow cells. After a period of free buffer flow, any analyteremaining bound to the surface may be removed with, for example, a pulseof 0.1% SDS bringing the signal back to baseline. Specific binding tothe derivatized sensor chips may be determined automatically by thesystem by subtraction of test from control flow cell responses. Ingeneral, OB-cadherin binds to an OB-cadherin extracellular domain (andN-cadherin binds to an N-cadherin extracellular domain) at a detectablelevel within such as assay.

[0036] OB-cadherin or N-cadherin activity may be evaluated using any ofa variety of in vitro assays designed to measure the effect on aresponse that is mediated by OB-cadherin or N-cadherin. The ability tomodulate cell adhesion may generally be evaluated in vitro by assayingthe effect on adhesion between cells expressing OB-cadherin orN-cadherin. In general, a polypeptide is an inhibitor of cell adhesionif contact of the test cells with the polypeptide results in adiscernible disruption of cell adhesion. Such assays may be performedusing any type of cell that expresses OB-cadherin or N-cadherin at adetectable level, using standard techniques such as immunocytochemicalprotocols (e.g., Blaschuk and Farookhi, Dev. Biol. 136:564-567, 1989).For example, such cells may be plated under standard conditions that, inthe absence of soluble OB-cadherin or N-cadherin, permit cell adhesion.In the presence of OB-cadherin or N-cadherin (e.g., 1 mg/mL), disruptionof cell adhesion may be determined visually within 24 hours, byobserving retraction of the cells from one another and the substratum.

[0037] Cells that express OB-cadherin include stromal, osteoblast andmetastatic cancer cells. Cells that express N-cadherin include neuralcells, endothelial cells and a variety of cancer cell types.Alternatively, cells that do not naturally express OB-cadherin orN-cadherin may be used within such assays. Such cells may be stablytransfected with a polynucleotide (e.g., a cDNA) encoding OB-cadherin orN-cadherin, such that OB-cadherin or N-cadherin is expressed on thesurface of the cell. Expression of OB-cadherin or N-cadherin may beconfirmed by assessing adhesion of the transfected cells, in conjunctionwith immunocytochemical techniques using antibodies directed againstOB-cadherin or N-cadherin. The stably transfected cells that aggregate,as judged by light microscopy, following transfection express sufficientlevels of OB-cadherin or N-cadherin. Preferred cells for use in suchassays include L cells, which do not detectably adhere and do notexpress any cadherin (Nagafuchi et al., Nature 329:341-343, 1987).Following transfection of L cells with a cDNA encoding OB-cadherin orN-cadherin, aggregation is observed (see Okazaki et al., J. Biol. Chem.269:12092-98, 1994). Peptides that detectably inhibit such aggregationmay be used to generate or characterize binding agents as describedherein.

[0038] By way of example, an assay for evaluating the ability to inhibitan OB-cadherin mediated function may employ MDA-231 human breast cancercells. According to a representative procedure, the cells may be platedat 10-20,000 cells per 35 mm tissue culture flasks containing DMEM with5% FCS and sub-cultured periodically (Sommers et al., Cell Growth Diffn2:365-72, 1991). Cells may be harvested and replated in 35 mm tissueculture flasks containing 1 mm coverslips and incubated until 50-65%confluent (24-36 hours). At this time, coverslips may be transferred toa 24-well plate, washed once with fresh DMEM and exposed to a testcompound (e g., protein or peptide) at a concentration of, for example,1 mg/mL for 24 hours. Fresh test compound may then be added, and thecells left for an additional 24 hours. Cells may be fixed with 2%paraformaldehyde for 30 minutes and then washed three times with PBS.Coverslips can be mounted and viewed by phase contrast microscopy.

[0039] In the absence of OB-cadherin or a portion thereof comprising acell adhesion recognition sequence, MDA-231 cells display anepithelial-like morphology and are well attached to the substratum.MDA-231 cells that are treated with such a peptide or protein may assumea round shape and become loosely attached to the substratum within 48hours of treatment with 1 mg/mL of OB-cadherin.

[0040] Alternatively, a protein or peptide may be assessed for theability to enhance skin permeability. This ability may be assessed bydetermining, for example, the effect on permeability of adherentepithelial and/or endothelial cell layers (e.g., human skin). Such skinmay be derived from a natural source or may be synthetic. Humanabdominal skin for use in such assays may generally be obtained fromhumans at autopsy within 24 hours of death. Briefly, a test compound(e.g., 500 μg/ml) and a marker (e.g., the fluorescent markers OregonGreen™ and Rhodamine Green™ Dextran) may be dissolved in a sterilebuffer (e.g., phosphate buffer, pH 7.2), and the ability of the markerto penetrate through the skin and into a receptor fluid (e.g., phosphatebuffer) may be measured using a Franz Cell apparatus (Franz, Curr. Prob.Dermatol. 7:58-68, 1978; Franz, J. Invest. Dermatol. 64:190-195, 1975).The penetration of the markers through the skin may be assessed at, forexample, 6, 12, 24, 36, and 48 hours after the start of the experiment.In general, OB-cadherin, N-cadherin or a portion thereof comprising acell adhesion recognition sequence should result in a statisticallysignificant increase in the amount of marker in the receptor compartmentafter 648 hours in the presence of 500 μg/mL peptide.

[0041] Yet another assay evaluates the ability to inhibit angiogenesis.The effect on angiogenesis may generally be determined by evaluating theeffect on blood vessel formation. Such a determination may generally beperformed, for example, using a chick chorioallantoic membrane assay(Iruela-Arispe et al., Molecular Biology of the Cell 6:327-343, 1995).Briefly, a peptide or protein may be embedded in a mesh composed ofvitrogen at one or more concentrations (e.g., ranging from about 1 to100 μg/mesh). The mesh(es) may then be applied to chick chorioallantoicmembranes. After 24 hours, the effect may be determined using computerassisted morphometric analysis. OB-cadherin, N-cadherin or a portionthereof comprising a CAR sequence should inhibit angiogenesis by atleast 25% at a concentration of 33 μg/mesh.

[0042] For N-cadherin, a neurite outgrowth assay may be used. Within arepresentative neurite outgrowth assay, neurons may be cultured on amonolayer of cells (e.g., 3T3) that express N-cadherin. Neurons grown onsuch cells (under suitable conditions and for a sufficient period oftime) extend longer neurites than neurons cultured on cells that do notexpress N-cadherin. For example, neurons may be cultured on monolayersof 3T3 cells transfected with cDNA encoding N-cadherin essentially asdescribed by Doherty and Walsh, Curr. Op. Neurobiol. 4:49-55, 1994;Williams et al., Neuron 13:583-594, 1994; Hall et al., Cell Adhesion andCommun. 3:441-450, 1996; Doherty and Walsh, Mol. Cell. Neurosci.8:99-111, 1994; and Safell et al., Neuron 18:231-242, 1997. Briefly,monolayers of control 3T3 fibroblasts and 3T3 fibroblasts that expressN-cadherin may be established by overnight culture of 80,000 cells inindividual wells of an 8-chamber well tissue culture slide. 3000cerebellar neurons isolated from post-natal day 3 mouse brains may becultured for 18 hours on the various monolayers in control media(SATO/2% FCS), or media supplemented with various concentrations of themodulating agent or control peptide. The cultures may then be fixed andstained for GAP43 which specifically binds to the neurons and theirneurites. The length of the longest neurite on each GAP43 positiveneuron may be measured by computer assisted morphometry. N-cadherin or aportion thereof comprising a cell adhesion recognition sequence (e.g.,500 μg/mL) should result in a decrease in the mean neurite length by atleast 50%, relative to the length in the absence of such a polypeptide.

[0043] Binding Agents

[0044] The term “binding agent” refers to a substance such as apolypeptide comprising an OB-cadherin or N-cadherin CAR sequence, orantibody (or antigen-binding fragment thereof) that specifically bindsto an OB-cadherin or N-cadherin. Certain preferred binding agents bindto an OB-cadherin or N-cadherin extracellular domain. As used herein, asubstance is said to “specifically bind” to an OB-cadherin or N-cadherinsequence if it reacts at a detectable level with OB-cadherin orN-cadherin, and does not react detectably with peptides containing anunrelated sequence or a sequence of a different cadherin. Such bindingproperties may generally be assessed using an ELISA, which may bereadily performed by those of ordinary skill in the art and isdescribed, for example, by Newton et al., Develop. Dynamics 197:1-13,1993. Each binding agent should satisfy the above criteria; however,those of ordinary skill in the art will recognize that binding agentsmay be used in combination to improve sensitivity. Peptides comprisingOB-cadherin or N-cadherin sequences for use in preparing and evaluatingbinding agents may be generated as described herein.

[0045] Any compound that satisfies the above requirements may be abinding agent. For example, a binding agent may be a ribosome, with orwithout a peptide component, an RNA molecule or a polypeptide. Withincertain preferred embodiments, a binding agent is a polypeptide thatcomprises an OB-cadherin or N-cadherin CAR sequence, a peptide variantthereof or a non-peptide mimetic of such a CAR sequence. As used herein,an “OB-cadherin CAR sequence” is a peptide portion of an OB-cadherinthat is capable of modulating a function mediated by OB-cadherin.Similarly, an “N-cadherin CAR sequence” is a peptide portion of aN-cadherin that is capable of modulating a function mediated byN-cadherin. A CAR sequence may be of any length, but generally comprisesat least three amino acid residues, preferably 4-16 amino acid residuesand more preferably 5-9 amino acid residues. An N-cadherin CAR sequencegenerally comprises the peptide His-Ala-Val (HAV), with or withoutadditional flanking sequence, which may be derived from theextracellular domain sequence provided in FIG. 3. Certain OB-cadherinCAR sequences have at least three consecutive amino acids present withinthe following consensus sequence in an OB-cadherin:Aaa-Phe-Val/Ser-Ile/Val-Asp/Glu-Baa- (SEQ ID NO:10) Caa-Ser/Thr-Gly

[0046] wherein Aaa, Baa and Caa are independently selected amino acidresidues; Val/Ser is an amino acid that is valine or serine; Ile/Val isan amino acid that is isoleucine or valine; Asp/Glu is an amino acidthat is aspartate or glutamate; and Ser/Thr is an amino acid that isserine or threonine. Representative OB-cadherin CAR sequences include:DDK, IDDK (SEQ ID NO:11) DDKS (SEQ ID NO:12), VIDDK (SEQ ID NO:13),IDDKS (SEQ ID NO:14), VIDDKS (SEQ ID NO:15), DDKSG (SEQ ID NO:16),IDDKSG (SEQ ID NO:17), VIDDKSG (SEQ ID NO:18), FVIDDK (SEQ ID NO:19),FVIDDKS (SEQ ID NO:20), FVIDDKSG (SEQ ID NO:21), IFVIDDK (SEQ ID NO:22),IFVIDDKS (SEQ ID NO:23), IFVIDDKSG (SEQ ID NO:24), EEY, IEEY (SEQ IDNO:25), EEYT (SEQ ID NO:26), VIEEY (SEQ ID NO:27), IEEYT (SEQ ID NO:28),VIEEYT (SEQ ID NO:29), EEYTG (SEQ ID NO:30), IEEYTG (SEQ ID NO:31),VIEEYTG (SEQ ID NO:32), FVIEEY (SEQ ID NO:33), FVIEEYT (SEQ ID NO:34),FVIEEYTG (SEQ ID NO:35), FFVIEEY (SEQ ID NO:36), FFVIEEYT (SEQ IDNO:37), FFVIEEYTG (SEQ ID NO:38), EAQ, VEAQ (SEQ ID NO:39), EAQT (SEQ IDNO:40), SVEAQ (SEQ ID NO:41), VEAQT (SEQ ID NO:42), SVEAQT (SEQ IDNO:43), EAQTG (SEQ ID NO:44), VEAQTG (SEQ ID NO:45), SVEAQTG (SEQ IDNO:46), FSVEAQ (SEQ ID NO:47), FSVEAQT (SEQ ID NO:48), FSVEAQTG (SEQ IDNO:49), YFSVEAQ (SEQ ID NO:50), YFSVEAQT (SEQ ID NO:51) or YFSVEAQTG(SEQ ID NO:52). Linear peptides having such sequences may be modified atthe N- and/or C-termini, as in the peptides N-Ac-IFVIDDKSG-NH₂ (SEQ IDNO:9), N-Ac-FFVIEEYTG-NH₂ (SEQ ID NO:53) and N-Ac-YFSVEAQTG-NH₂ (SEQ IDNO:54).

[0047] To confirm that a particular sequence is an OB-cadherin orN-cadherin CAR sequence, the ability of the sequence to modulate afunction mediated by an OB-cadherin may be determined. For functions(e.g., cell adhesion) that are inhibited by a full length OB-cadherin orN-cadherin, a CAR sequence should inhibit the function with an activitythat is not substantially diminished relative to the full lengthOB-cadherin or N-cadherin (i.e., the CAR sequence inhibits the functionat least as well as soluble OB-cadherin or N-cadherin, when contactedwith cells that express OB-cadherin or N-cadherin).

[0048] The present invention further contemplates the use of OB-cadherinand N-cadherin CAR sequences from other organisms. Such CAR sequencesmay be identified based upon similarity to the sequences providedherein, and the ability to modulate an OB-cadherin- orN-cadherin-mediated function may be confirmed as described herein.

[0049] As noted above, binding agents as described herein may comprisean analogue or mimetic of an OB-cadherin or N-cadherin CAR sequence. Ananalogue generally retains at least 50% identity to a native OB-cadherinor N-cadherin CAR sequence, and modulates an OB-cadherin- orN-cadherin-mediated function as described herein. Such analoguespreferably contain at least three consecutive residues of, and morepreferably at least five consecutive residues of, an OB-cadherin orN-cadherin CAR sequence. An analogue may contain any of a variety ofamino acid substitutions, additions, deletions and/or modifications(e.g., side chain modifications). Preferred amino acid substitutions areconservative. A “conservative substitution” is one in which an aminoacid is substituted for another amino acid that has similar properties,such that one skilled in the art of peptide chemistry would expect thesecondary structure and hydropathic nature of the polypeptide to besubstantially unchanged. Amino acid substitutions may generally be madeon the basis of similarity in polarity, charge, solubility,hydrophobicity, hydrophilicity and/or the amphipathic nature of theresidues. For example, negatively charged amino acids include asparticacid and glutamic acid; positively charged amino acids include lysineand arginine; and amino acids with uncharged polar head groups havingsimilar hydrophilicity values include leucine, isoleucine and valine;glycine and alanine; asparagine and glutamine; and serine, threonine,phenylalanine and tyrosine. Other groups of amino acids that mayrepresent conservative changes include: (1) ala, pro, gly, glu, asp,gln, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala,phe; (4) lys, arg, his; and (5) phe, tyr, trp, his. The criticaldetermining feature of an OB-cadherin or N-cadherin CAR sequenceanalogue is the ability to modulate an OB-cadherin- orN-cadherin-mediated function, which may be evaluated using therepresentative assays provided herein.

[0050] A mimetic is a non-peptidyl compound that is conformationallysimilar to an OB-cadherin or N-cadherin CAR sequence, such that itmodulates an OB-cadherin- or N-cadherin-mediated function as describedherein. Such mimetics may be designed based on techniques that evaluatethe three dimensional structure of a peptide. For example, NuclearMagnetic Resonance spectroscopy (NMR) and computational techniques maybe used to determine the conformation of a CAR sequence. NMR is widelyused for structural analyses of both peptidyl and non-peptidylcompounds. Nuclear Overhauser Enhancements (NOE's), coupling constantsand chemical shifts depend on the conformation of a compound. NOE dataprovides the interproton distance between protons through space and canbe used to calculate the lowest energy conformation for the CARsequence. This information can then be used to design mimetics of thepreferred conformation. Linear peptides in solution exist in manyconformations. By using conformational restriction techniques it ispossible to fix the peptide in the active conformation. Conformationalrestriction can be achieved by i) introduction of an alkyl group such asa methyl which sterically restricts free bond rotation; ii) introductionof unsaturation which fixes the relative positions of the terminal andgeminal substituents; and/or iii) cyclization, which fixes the relativepositions of the sidechains. Mimetics may be synthesized where one ormore of the amide linkages has been replaced by isosteres, substituentsor groups which have the same size or volume such as —CH₂NH—, —CSNH—,—CH₂S—, —CH═CH—, —CH₂CH₂—, —CONMe— and others. These backbone amidelinkages can also be part of a ring structure (e.g., lactam). Mimeticsmay be designed where one or more of the side chain functionalities of aCAR sequence are replaced by groups that do not necessarily have thesame size or volume, but have similar chemical and/or physicalproperties which produce similar biological responses. Other mimeticsmay be small molecule mimics, which may be readily identified from smallmolecule libraries, based on the three-dimensional structure of a CARsequence. It should be understood that, within embodiments describedbelow, an analogue or mimetic may be substituted for an OB-cadherin orN-cadherin CAR sequence.

[0051] Binding agents, or peptide portions thereof, may be linear orcyclic peptides. The term “cyclic peptide,” as used herein, refers to apeptide or salt thereof that comprises (I) an intramolecular covalentbond between two non-adjacent residues and (2) at least one OB-cadherinor N-cadherin CAR sequence or an analogue thereof. The intramolecularbond may be a backbone to backbone, side-chain to backbone or side-chainto side-chain bond (ie., terminal functional groups of a linear peptideand/or side chain functional groups of a terminal or interior residuemay be linked to achieve cyclization). Preferred intramolecular bondsinclude, but are not limited to, disulfide, amide and thioether bonds.

[0052] The size of a cyclic peptide ring generally ranges from 5 toabout 15 residues, preferably from 5 to 10 residues. Additionalresidue(s) may be present on the N-terminal and/or C-terminal side of anOB-cadherin or N-cadherin CAR sequence, and may be derived fromsequences that flank an OB-cadherin or N-cadherin CAR sequence, with orwithout amino acid substitutions and/or other modifications.Alternatively, additional residues present on one or both sides of theCAR sequence(s) may be unrelated to an endogenous sequence (e.g.,residues that facilitate cyclization, purification or other manipulationand/or residues having a targeting or other function).

[0053] Within certain embodiments, a binding agent may comprise a cyclicpeptide having the formula:

[0054] Within this formula, W is a tripeptide selected from the groupconsisting of EEY, DDK and EAQ (for OB-cadherin) or HAV (forN-cadherin); X₁, and X₂ are optional, and if present, are independentlyselected from the group consisting of amino acid residues andcombinations thereof in which the residues are linked by peptide bonds,and wherein X₁ and X₂ independently range in size from 0 to 10 residues,such that the sum of residues contained within X₁ and X₂ ranges from 1to 12; Y₁ and Y₂ are independently selected from the group consisting ofamino acid residues, and wherein a covalent bond is formed betweenresidues Y₁ and Y₂; and Z₁ and Z₂ are optional, and if present, areindependently selected from the group consisting of amino acid residuesand combinations thereof in which the residues are linked by peptidebonds.

[0055] For example, representative cyclic peptides comprising anOB-cadherin CAR sequence include CDDKC (SEQ ID NO:55), CIDDKC (SEQ IDNO:56), CDDKSC (SEQ ID NO:57), CVIDDKC (SEQ ID NO:58), CIDDKSC (SEQ IDNO:59), CVIDDKSC (SEQ ID NO:60), CDDKSGC (SEQ ID NO:61), CIDDKSGC (SEQID NO:62), CVIDDKSGC (SEQ ID NO:63), CFVIDDKC (SEQ ID NO:64), CFVIDDKSC(SEQ ID NO:65), CFVIDDKSGC (SEQ ID NO:66), CIFVIDDKC (SEQ ID NO:67),CIFVIDDKSC (SEQ ID NO:68), CIFVIDDKSGC (SEQ ID NO:69), DDDKK (SEQ IDNO:70), DIDDKK (SEQ ID NO:71), DVIDDKK (SEQ ID NO:72), DFVIDDKK (SEQ IDNO:73), DIFVIDDKK (SEQ ID NO:74), EDDKK (SEQ ID NO:75), EIDDKK (SEQ IDNO:76), EVIDDKK (SEQ ID NO:77), EFVIDDKK (SEQ ID NO:78), EIFVIDDKK (SEQID NO:79), FVIDDK (SEQ ID NO:80), FVIDDKS (SEQ ID NO:81), FVIDDKSG (SEQID NO:82), KDDKD (SEQ ID NO:83), KIDDKD (SEQ ID NO:84), KDDKSD (SEQ IDNO:85), KVIDDKD (SEQ ID NO:86), KIDDKSD (SEQ ID NO:87), KVIDDKSD (SEQ IDNO:88), KDDKSGD (SEQ ID NO:89), KIDDKSGD (SEQ ID NO:90), KVIDDKSGD (SEQID NO:91), KFVIDDKD (SEQ ID NO:92), KFVIDDKSD (SEQ ID NO:93), KFVIDDKSGD(SEQ ID NO:94), KIFVIDDKD (SEQ ID NO:95), KIFVIDDKSD (SEQ ID NO:96),KIFVIDDKSGD (SEQ ID NO:97), VIDDK (SEQ ID NO:98), IDDKS (SEQ ID NO:99),VIDDKS (SEQ ID NO:100), VIDDKSG (SEQ ID NO:101), DDKSG (SEQ ID NO:102),IDDKSG (SEQ ID NO:103), IFVIDDK (SEQ ID NO:104), IFVIDDKS (SEQ IDNO:105), IFVIDDKSG (SEQ ID NO:106), KDDKE (SEQ ID NO:107), KIDDKE (SEQID NO:108), KDDKSE (SEQ ID NO:109), KVIDDKE (SEQ ID NO:110), KIDDKSE(SEQ ID NO:111), KVIDDKSE (SEQ ID NO:112), KDDKSGE (SEQ ID NO:113),KIDDKSGE (SEQ ID NO:114), KVIDDKSGE (SEQ ID NO:115), KFVIDDKE (SEQ IDNO:116), KFVIDDKSE (SEQ ID NO:117), KFVIDDKSGE (SEQ ID NO:118),KIFVIDDKE (SEQ ID NO:119), KIFVIDDKSE (SEQ ID NO:120), KIFVIDDKSGE (SEQID NO:121), CEEYC (SEQ ID NO:122), CIEEYC (SEQ ID NO:123), CEEYTC (SEQID NO:124), CVIEEYC (SEQ ID NO:125), CIEEYTC (SEQ ID NO:126), CVIEEYTC(SEQ ID NO:127), CEEYTGC (SEQ ID NO:128), CIEEYTGC (SEQ ID NO:129),CVIEEYTGC (SEQ ID NO:130), CFVIEEYC (SEQ ID NO:131), CFVIEEYTC (SEQ IDNO:132), CFVIEEYTGC (SEQ ID NO:133), CFFVIEEYC (SEQ ID NO:134),CFFVIEEYTC (SEQ ID NO:135), CFFVIEEYTGC (SEQ ID NO:136), KEEYD (SEQ IDNO:137), KIEEYD (SEQ ID NO:138), KEEYTD (SEQ ID NO:139), KVIEEYD (SEQ IDNO:140), KIEEYTD (SEQ ID NO:141), KVIEEYTD (SEQ ID NO:142), KEEYTGCD(SEQ ID NO:143), KIEEYTGD (SEQ ID NO:144), KVIEEYTGD (SEQ ID NO:145),KFVIEEYD (SEQ ID NO:146), KFVIEEYTD (SEQ ID NO:147), KFVIEEYTGD (SEQ IDNO:148), KFFVIEEYD (SEQ ID NO:149), KFFVIEEYTD (SEQ ID NO:150),KFFVIEEYTGD (SEQ ID NO:151), EEEYK (SEQ ID NO:152), EIEEYK (SEQ IDNO:153), EEEYTK (SEQ ID NO:154), EVIEEYK (SEQ ID NO:155), EIEEYTK (SEQID NO:156), EVIEEYTK (SEQ ID NO:157), EEEYTGK (SEQ ID NO:158), EIEEYTGK(SEQ ID NO:159), EVIEEYTGK (SEQ ID NO:160), EFVIEEYK (SEQ ID NO:161),EFVIEEYTK (SEQ ID NO:162), EFVIEEYTGK (SEQ ID NO:163), EFFVIEEYK (SEQ IDNO:164), EFFVIEEYTK (SEQ ID NO:165), EFFVIEEYTGK (SEQ ID NO:166), DCEEYK(SEQ ID NO:167), DIEEYCK (SEQ ID NO:168), DEEYTK (SEQ ID NO:169),DVIEEYK (SEQ ID NO:170), DIEEYTK (SEQ ID NO:171), DVIEEYTK (SEQ IDNO:172), DEEYTGK (SEQ ID NO:173), DIEEYTGK (SEQ ID NO:174), DVIEEYTGK(SEQ ID NO:175), DFVIEEYK (SEQ ID NO:176), DFVIEEYTK (SEQ ID NO:177),DFVIEEYTGK (SEQ ID NO:178), DFFVIEEYK (SEQ ID NO:179), DFFVIEEYTK (SEQID NO:180), DFFVIEEYTGK (SEQ ID NO:181), KEEYE (SEQ ID NO:182), KIEEYE(SEQ ID NO:183), KEEYTE (SEQ ID NO:184), KVIEEYE (SEQ ID NO:185),KIEEYTE (SEQ ID NO:186), KVIEEYTE (SEQ ID NO:187), KEEYTGE (SEQ IDNO:188), KIEEYTGE (SEQ ID NO:189), KVIEEYTGE (SEQ ID NO:190), KFVIEEYE(SEQ ID NO:191), KFVIEEYTE (SEQ ID NO:192), KFVIEEYTGE (SEQ ID NO:193),KFFVIEEYE (SEQ ID NO:194), KFFVIEEYTE (SEQ ID NO:195), KFFVIEEYTGE (SEQID NO:196), VIEEY (SEQ ID NO:197), IEEYT (SEQ ID NO:198), VIEEYT (SEQ IDNO:199), EEYTG (SEQ ID NO:200), IEEYTG (SEQ ID NO:201), VIEEYTG (SEQ IDNO:202), FVIEEY (SEQ ID NO:203), FVIEEYT (SEQ ID NO:204), FVIEEYTG (SEQID NO:205), FFVIEEY (SEQ ID NO:206), FFVIEEYT (SEQ ID NO:207), FFVIEEYTG(SEQ ID NO:208), CEAQC (SEQ ID NO:209), CVEAQC (SEQ ID NO:210), CEAQTC(SEQ ID NO:211), CSVEAQC (SEQ ID NO:212), CVEAQTC (SEQ ID NO:213),CSVEAQTC (SEQ ID NO:214), CEAQTGC (SEQ ID NO:215), CVEAQTGC (SEQ IDNO:216), CSVEAQTGC (SEQ ID NO:217), CFSVEAQC (SEQ ID NO:218), CFSVEAQTC(SEQ ID NO:219), CFSVEAQTGC (SEQ ID NO:220), CYFSVEAQC (SEQ ID NO:221),CYFSVEAQTC (SEQ ID NO:222), CYFSVEAQTGC (SEQ ID NO:223), KEAQD (SEQ IDNO:224), KVEAQD (SEQ ID NO:225), KEAQTD (SEQ ID NO:226), KSVEAQD (SEQ IDNO:227), KVEAQTD (SEQ ID NO:228), KSVEAQTD (SEQ ID NO:229), KEAQTGD (SEQID NO:230), KVEAQTGD (SEQ ID NO:231), KSVEAQTGD (SEQ ID NO:232),KFSVEAQD (SEQ ID NO:233), KFSVEAQTD (SEQ ID NO:234), KFSVEAQTGD (SEQ IDNO:235), KYFSVEAQD (SEQ ID NO:236), KYFSVEAOTD (SEQ ID NO:237),KYFSVEAQTGD (SEQ ID NO:238), EEAQK (SEQ ID NO:239), EVEAQK (SEQ IDNO:240), EEAQTK (SEQ ID NO:241), ESVEAQK (SEQ ID NO:242), EVEAQTK (SEQID NO:243), ESVEAQTK (SEQ ID NO:244), EEAQTGK (SEQ ID NO:245), EVEAQTGK(SEQ ID NO:246), ESVEAQTGK (SEQ ID NO:247), EFSVEAQK (SEQ ID NO:248),EFSVEAQTK (SEQ ID NO:249), EFSVEAQTGK (SEQ ID NO:250), EYFSVEAQK (SEQ IDNO:251), EYFSVEAQTK (SEQ ID NO:252), EYFSVEAQTGK (SEQ ID NO:253), DEAQK(SEQ ID NO:254), DVEAQK (SEQ ID NO:255), DEAQTK (SEQ ID NO:256), DSVEAQK(SEQ ID NO:257), DVEAQTK (SEQ ID NO:258), DSVEAQTK (SEQ ID NO:259),DEAQTGK (SEQ ID NO:260), DVEAQTGK (SEQ ID NO:261), DSVEAQTGK (SEQ IDNO:262), DFSVEAQK (SEQ ID NO:263), DFSVEAQTK (SEQ ID NO:264), DFSVEAQTGK(SEQ ID NO:265), DYFSVEAQK (SEQ ID NO:266), DYFSVEAQTK (SEQ ID NO:267),DYFSVEAQTGK (SEQ ID NO:268), KEAQE (SEQ ID NO:269), KVEAQE (SEQ IDNO:270), KEAQTE (SEQ ID NO:271), KSVEAQE (SEQ ID NO:272), KVEAQTE (SEQID NO:273), KSVEAQTE (SEQ ID NO:274), KEAQTGE (SEQ ID NO:275), KVEAQTGE(SEQ ID NO:276), KSVEAQTGE (SEQ ID NO:277), KFSVEAQE (SEQ ID NO:278),KFSVEAQTE (SEQ ID NO:279), KFSVEAQTGE (SEQ ID NO:280), KYFSVEAQE (SEQ IDNO:281), KYFSVEAQTE (SEQ ID NO:282), KYFSVEAQTGE (SEQ ID NO:283), SVEAQ(SEQ ID NO:284), VEAQT (SEQ ID NO:285), SVEAQT (SEQ ID NO:286), EAQTG(SEQ ID NO:287), VEAQTG (SEQ ID NO:288), SVEAQTG (SEQ ID NO:289), FSVEAQ(SEQ ID NO:290), FSVEAQT (SEQ ID NO:291), FSVEAQTG (SEQ ID NO:292),YFSVEAQ (SEQ ID NO:293), YFSVEAQT (SEQ ID NO:294) and YFSVEAQTG (SEQ IDNO:295). Within the context of the present invention, underlinedsequences are cyclized using any suitable method, as described herein.

[0056] Representative cyclic peptides comprising an N-cadherin CARsequence include CHAVC (SEQ ID NO:296), CHAVDC (SEQ ID NO:297), CAHAVC(SEQ ID NO:298), CAHAVDC (SEQ ID NO:299), CAHAVDIC (SEQ ID NO:300),CRAHAVDC (SEQ ID NO:301), CLRAHAVC (SEQ ID NO:302), CLRAHAVDC (SEQ IDNO:303), CSHAVC (SEQ ID NO:304), CHAVSC (SEQ ID NO:305), CSHAVSC (SEQ IDNO:306), CSHAVSSC (SEQ ID NO:307), CHAVSSC (SEQ ID NO:308), KHAVD (SEQID NO:309), DHAVK (SEQ ID NO:310), KHAVE (SEQ ID NO:311), AHAVDI (SEQ IDNO:312), SHAVDSS (SEQ ID NO:313) and KSHAVSSD (SEQ ID NO:314).

[0057] In certain other preferred embodiments, a binding agent is anantibody or an antigen-binding fragment thereof. Such antibodies may bepolyclonal or monoclonal. In addition, the antibodies may be singlechain, chimeric, CDR-grafted or humanized.

[0058] Polyclonal and monoclonal antibodies may be raised against anOB-cadherin or N-cadherin sequence using conventional techniques. See,e.g., Harlow and Lane, Antibodies: A Laboratory Manual, Cold SpringHarbor Laboratory, 1988. In one such technique, an immunogen comprisingthe sequence is initially injected into any of a wide variety of mammals(e.g., mice, rats, rabbits, sheep or goats). The smaller immunogens(i.e., less than about 20 amino acids) should be joined to a carrierprotein, such as bovine serum albumin or keyhole limpet hemocyanin.Following one or more injections, the animals are bled periodically.Polyclonal antibodies specific for the OB-cadherin or N-cadherinsequence may then be purified from such antisera by, for example,affinity chromatography using the OB-cadherin or N-cadherin sequence orantigenic portion thereof coupled to a suitable solid support.

[0059] Monoclonal antibodies specific for an OB-cadherin or N-cadherinsequence may be prepared, for example, using the technique of Kohler andMilstein, Eur. J. Immunol. 6:511-519, 1976, and improvements thereto.Briefly, these methods involve the preparation of immortal cell linescapable of producing antibodies having the desired specificity fromspleen cells obtained from an animal immunized as described above. Thespleen cells are immortalized by, for example, fusion with a myelomacell fusion partner, preferably one that is syngeneic with the immunizedanimal. Single colonies are selected and their culture supernatantstested for binding activity against the OB-cadherin or N-cadherinsequence or antigenic portion thereof. Hybridomas having high reactivityand specificity are preferred.

[0060] Monoclonal antibodies may be isolated from the supernatants ofgrowing hybridoma colonies, with or without the use of varioustechniques known in the art to enhance the yield. Contaminants may beremoved from the antibodies by conventional techniques, such aschromatography, gel filtration, precipitation and extraction. Antibodieshaving the desired activity may generally be identified usingimmunofluorescence analyses of tissue sections, cell or other sampleswhere the target is cadherin is localized.

[0061] Within certain embodiments, the use of antigen-binding fragmentsof antibodies may be preferred. Such fragments include Fab fragments,which may be prepared using standard techniques. Briefly,immunoglobulins may be purified from rabbit serum by affinitychromatography on Protein A bead columns (Harlow and Lane, Antibodies: ALaboratory Manual, Cold Spring Harbor Laboratory, 1988; see especiallypage 309) and digested by papain to yield Fab and Fc fragments. The Faband Fc fragments may be separated by affinity chromatography on proteinA bead columns (Harlow and Lane, 1988, pages 628-29).

[0062] OB-Cadherin and N-Cadherin Polynucleotides

[0063] A polynucleotide that encodes OB-cadherin or N-cadherin (or aportion or other variant thereof) or that is complementary to such apolynucleotide, may be used within certain methods provided herein.Polynucleotides may be single-stranded (coding or antisense) ordouble-stranded, and may be DNA (cDNA or synthetic) or RNA molecules.Additional coding or non-coding sequences may, but need not, be presentwithin a polynucleotide of the present invention, and a polynucleotidemay, but need not, be linked to other molecules and/or supportmaterials. Oligonucleotides for use within the methods provided hereinmay be of any length suitable for the particular assay.

[0064] Polynucleotides may comprise a native sequence (i.e., anendogenous sequence that encodes OB-cadherin, N-cadherin or a portionthereof) or may comprise a variant of such a sequence. Polynucleotidevariants preferably exhibit at least about 70% identity, more preferablyat least about 80% identity and most preferably at least about 90%identity to a polynucleotide sequence that encodes a native OB-cadherin,N-cadherin or a portion thereof. Certain variants are substantiallyhomologous to a native gene, or a portion or complement thereof. Suchpolynucleotide variants are capable of hybridizing under moderatelystringent conditions to a naturally occurring DNA sequence encoding anative OB-cadherin or N-cadherin (or a complementary sequence). Suitablemoderately stringent conditions include prewashing in a solution of5×SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50° C.-65° C.,5×SSC, overnight; followed by washing twice at 65° C. for 20 minuteswith each of 2×, 0.5× and 0.2×SSC containing 0.1% SDS.

[0065] Polynucleotides may be prepared using any of a variety oftechniques. For example, a polynucleotide may be amplified viapolymerase chain reaction (PCR) from cDNA prepared from cells expressingOB-cadherin or N-cadherin. For this approach, sequence-specific primersmay be designed based on the sequences provided herein, and may bepurchased or synthesized. Other polynucleotides may be directlysynthesized by any method known in the art, including chemical synthesisby, for example, solid phase phosphoramidite chemical synthesis.Modifications in a polynucleotide sequence may also be introduced usingstandard mutagenesis techniques, such as oligonucleotide-directedsite-specific mutagenesis. RNA molecules may be generated by in vitro orin vivo transcription of DNA sequences encoding OB-cadherin orN-cadherin, or portion thereof, provided that the DNA is incorporatedinto a vector with a suitable RNA polymerase promoter (such as T7 orSP6).

[0066] Particularly preferred portions of a coding sequence or acomplementary sequence are those designed as a probe or primer to detectgene expression. Probes may be labeled by a variety of reporter groups,such as radionuclides and enzymes, and are preferably at least 10nucleotides in length, more preferably at least 20 nucleotides in lengthand still more preferably at least 30 nucleotides in length. Primers arepreferably 22-30 nucleotides in length.

[0067] Any polynucleotide may be further modified to increase stabilityin vivo. Possible modifications include, but are not limited to, theaddition of flanking sequences at the 5′ and/or 3′ ends; the use ofphosphorothioate or 2′ O-methyl rather than phosphodiesterase linkagesin the backbone; and/or the inclusion of nontraditional bases such asinosine, queosine and wybutosine, as well as acetyl- methyl-, thio- andother modified forms of adenine, cytidine, guanine, thymine and uridine.

[0068] Nucleotide sequences as described herein may be joined to avariety of other nucleotide sequences using established recombinant DNAtechniques. For example, a polynucleotide may be cloned into any of avariety of cloning vectors, including plasmids, phagemids, lambda phagederivatives and cosmids. Vectors of particular interest includeexpression vectors, replication vectors, probe generation vectors andsequencing vectors. In general, a vector will contain an origin ofreplication functional in at least one organism, convenient restrictionendonuclease sites and one or more selectable markers. Other elementswill depend upon the desired use, and will be apparent to those ofordinary skill in the art.

[0069] Methods for Diagnosing Cancer

[0070] Binding agents (such as OB-cadherin or N-cadherin CAR sequencesand antibodies raised against an OB-cadherin or N-cadherin sequence), aswell as polynucleotide probes and primers may be used for a variety ofdiagnostic and assay purposes. In general, such binding agents andpolynucleotides may be used to detect a metastatic cancer in a patient,to monitor progression of a cancer or to evaluate the metastaticpotential of a cancer. It has been found, within the context of thepresent invention, that OB-cadherin and/or N-cadherin are expressed byhighly invasive cancer cells. Such cells do not generally expressE-cadherin at a detectable level. In contrast, highly differentiated,poorly invasive carcinomas express E-cadherin, but do not expressOB-cadherin and/or N-cadherin. Accordingly, a metastatic cancer may bedetected in a patient based on an elevated level of OB-cadherin orN-cadherin (or RNA encoding OB-cadherin or N-cadherin) present within abiological sample obtained from the patient. Further informationregarding metastatic potential of a cancer may be obtained by alsoevaluating E-cadherin expression within the same or a similar sample.

[0071] Biological samples for use within such assays include blood,sera, urine, tumor or normal tissue biopsies, lymph node, peritonealfluid, cerebrospinal fluid and prostate secretions, as well as othertissues, homogenates, and extracts thereof. For assays employingpolynucleotide probes or primers, a biological sample may be a totalRNA, mRNA or cDNA preparation from any of the foregoing samples. Suchbiological samples may be prepared using any standard technique. Samplesmay be obtained from patients with or without a known cancer (asdetermined using standard clinical tests).

[0072] There are a variety of assay formats known to those of ordinaryskill in the art for using a binding agent to detect a target moleculein a sample. See, e.g., Harlow and Lane, Antibodies: A LaboratoryManual, Cold Spring Harbor Laboratory, 1988. In general, the presence orabsence of a cancer in a patient may be determined by (a) contacting abiological sample obtained from a patient with a binding agent; (b)detecting in the sample a level of polypeptide that binds to the bindingagent; and (c) comparing the level of polypeptide with a predeterminedcut-off value.

[0073] For example, an assay may be performed in a Western blot format,wherein a protein preparation from the biological sample is submitted togel electrophoresis, transferred to a suitable membrane and allowed toreact with an antibody binding agent. The antibody may be labeled, orthe presence of the antibody on the membrane may be detected using asuitable detection reagent, as described below. A similar assay may beperformed using a peptide binding agent comprising a CAR sequence. Suchbinding agents are generally labeled, as described below.

[0074] In a preferred embodiment, the assay involves the use of bindingagent immobilized on a solid support to bind to OB-cadherin orN-cadherin, or a proteolytic fragment thereof, and remove it from theremainder of the sample. The bound cadherin may then be detected using asecond binding agent or other detection reagent that contains a reportergroup and specifically binds to the binding agent/polypeptide complex.Such detection reagents may comprise, for example, a binding agent thatspecifically binds to OB-cadherin or N-cadherin. Alternatively, acompetitive assay may be utilized, in which OB-cadherin or N-cadherin,or a portion thereof, is labeled with a reporter group and allowed tobind to the immobilized binding agent after incubation of the bindingagent with the sample. The extent to which components of the sampleinhibit the binding of the labeled OB-cadherin or N-cadherin to thebinding agent is indicative of the reactivity of the sample with theimmobilized binding agent, and as a result, indicative of the level ofOB-cadherin or N-cadherin in the sample.

[0075] The solid support may be any material known to those of ordinaryskill in the art to which the binding agent may be attached, such as atest well in a microtiter plate, a nitrocellulose filter or anothersuitable membrane. Alternatively, the support may be a bead or disc,such as glass, fiberglass, latex or a plastic such as polystyrene orpolyvinylchloride. A binding agent, such as an antibody or peptide, maybe immobilized on the solid support using a variety of techniques knownto those in the art, which are amply described in the patent andscientific literature.

[0076] In certain embodiments, the assay for detection of OB-cadherin orN-cadherin in a sample is a two-antibody sandwich assay. This assay maybe performed by first contacting an antibody that has been immobilizedon a solid support, commonly the well of a microtiter plate, with thebiological sample, such that OB-cadherin or N-cadherin within the sampleis allowed to bind to the immobilized antibody (a 30 minute incubationtime at room temperature is generally sufficient). Unbound sample isthen removed from the immobilized cadherin-antibody complexes and asecond antibody (containing a reporter group such as an enzyme (such ashorseradish peroxidase), substrate, cofactor, inhibitor, dye,radionuclide, luminescent group, fluorescent group or biotin) capable ofbinding to a different site on the cadherin is added. The conjugation ofantibody to reporter group may be achieved using standard methods knownto those of ordinary skill in the art. The amount of second antibodythat remains bound to the solid support is then determined using amethod appropriate for the specific reporter group.

[0077] More specifically, once an antibody is immobilized on the supportas described above, the remaining protein binding sites on the supportare typically blocked. Any suitable blocking agent known to those ofordinary skill in the art, such as bovine serum albumin or Tween 20™(Sigma Chemical Co., St. Louis, Mo.). The immobilized antibody is thenincubated with the sample, and polypeptides within the sample areallowed to bind to the antibody. The sample may be diluted with asuitable diluent, such as phosphate-buffered saline (PBS) prior toincubation. In general, an appropriate contact time (i.e., incubationtime) is a period of time that is sufficient to detect the presence ofOB-cadherin or N-cadherin within a sample obtained from an individualwith a metastatic cancer. Preferably, the contact time is sufficient toachieve a level of binding that is at least about 95% of that achievedat equilibrium between bound and unbound polypeptide. Those of ordinaryskill in the art will recognize that the time necessary to achieveequilibrium may be readily determined by assaying the level of bindingthat occurs over a period of time. At room temperature, an incubationtime of about 30 minutes is generally sufficient.

[0078] Unbound sample may then be removed by washing the solid supportwith an appropriate buffer, such as PBS containing 0.1% Tween 20™. Thedetection reagent is then incubated with the immobilizedantibody-polypeptide complex for an amount of time sufficient to detectthe bound polypeptide. An appropriate amount of time may generally bedetermined by assaying the level of binding that occurs over a period oftime. Unbound detection reagent is then removed and bound detectionreagent is detected using the reporter group. The method employed fordetecting the reporter group depends upon the nature of the reportergroup. For radioactive groups, scintillation counting orautoradiographic methods are generally appropriate. Spectroscopicmethods may be used to detect dyes, luminescent groups and fluorescentgroups. Biotin may be detected using avidin, coupled to a differentreporter group (commonly a radioactive or fluorescent group or anenzyme). Enzyme reporter groups may generally be detected by theaddition of substrate (generally for a specific period of time),followed by spectroscopic or other analysis of the reaction products.Standards and standard additions may be used to determine the level ofOB-cadherin or N-cadherin in a sample, using well known techniques.

[0079] To determine the presence or absence of a metastatic cancer, thesignal detected from the reporter group that remains bound to the solidsupport is generally compared to a signal that corresponds to apredetermined cut-off value. In one preferred embodiment, the cut-offvalue for the detection of a metastatic cancer is the average meansignal obtained when the immobilized antibody is incubated with samplesfrom patients without a detectable cancer. In general, a samplegenerating a signal that is statistically greater (preferably two foldgreater) than the predetermined cut-off value is considered positive fora metastatic cancer. The precise cancer may be determined based onlocation of a tumor and/or using other clinically acceptable diagnostictechniques.

[0080] It will be apparent that numerous other assay protocols existthat are suitable for use with the antigens or binding agents of thepresent invention. For example, flow cytometry techniques may beapplied, as described by Seline et al., J. Invest. Dermatol.106:1320-1324, 1996. The above descriptions are intended to be exemplaryonly.

[0081] In another embodiment, binding agents may be used to monitor theprogression of a cancer. In this embodiment, assays as described abovefor the diagnosis of a metastatic cancer may be performed over time, andthe change in the level of reactive polypeptide(s) evaluated. Ingeneral, a cancer is progressing in those patients in whom the level ofpolypeptide detected by the binding agent increases over time. Incontrast, the cancer is not progressing when the level of reactivepolypeptide either remains constant or decreases with time.

[0082] Certain in vivo diagnostic assays may be performed directly on atumor. One such assay involves contacting tumor cells with a bindingagent. The bound binding agent may then be detected directly orindirectly via a reporter group. Such binding agents may also be used inhistological applications. Samples suitable for immunocytochemicalstaining of OB-cadherin may be prepared by any of a variety oftechniques. For example, frozen or paraffin-embedded tissue sections maybe prepared as described by Byers et al., Endocrinology 134:630-639,1994 and Cyr et al., Endocrinology 130:353-363, 1992, respectively, andplaced on glass slides. Alternatively, cells obtained from sources suchas peripheral blood or ascites fluid may be fixed onto glass slides asdescribed by Blaschuk and Farookhi, Dev. Biol. 136:564-567, 1989.Samples may then be probed using an anti-OB-cadherin antibody or labeledpeptide comprising a CAR sequence, using standard techniques.

[0083] As noted above, a metastatic cancer may also, or alternatively,be detected based on the level of mRNA encoding OB-cadherin orN-cadherin in a biological sample. For example, at least twooligonucleotide primers may be employed in a polymerase chain reaction(PCR) based assay to amplify a portion of OB-cadherin or N-cadherin cDNAderived from a biological sample, wherein at least one of theoligonucleotide primers is specific for (i.e., hybridizes to) apolynucleotide encoding OB-cadherin or N-cadherin. The amplified cDNA isthen separated and detected using techniques well known in the art, suchas gel electrophoresis. Similarly, oligonucleotide probes thatspecifically hybridize to a polynucleotide encoding OB-cadherin orN-cadherin may be used in a hybridization assay to detect the presenceof polynucleotide encoding the antigen in a biological sample.

[0084] To permit hybridization under assay conditions, oligonucleotideprimers and probes should comprise an oligonucleotide sequence that hasat least about 60%, preferably at least about 75% and more preferably atleast about 90%, identity to a portion of a polynucleotide encodingOB-cadherin or N-cadherin that is at least 10 nucleotides, andpreferably at least 20 nucleotides, in length. Oligonucleotide primersand/or probes which may be usefully employed in the diagnostic methodsdescribed herein preferably are at least 10-40 nucleotides in length. Ina preferred embodiment, the oligonucleotide primers comprise at least 10contiguous nucleotides, more preferably at least 15 contiguousnucleotides, of a DNA molecule encoding OB-cadherin. Techniques for bothPCR based assays and hybridization assays are well known in the art.

[0085] One preferred assay employs reverse transcriptase-polymerasechain reaction (RT-PCR), in which PCR is applied in conjunction withreverse transcription. Typically, RNA is extracted from a sample tissueusing standard techniques (e.g., guanidine isothiocyanate extraction asdescribed by Chomczynski and Sacchi, Anal. Biochem 162:156-159, 1987)and is reverse transcribed to produce cDNA molecules. This cDNA is thenused as a template for a subsequent polymerase chain reaction. The cDNAis hybridized to sets of primers, at least one of which is specificallydesigned against an OB-cadherin or N-cadherin sequence. Examples ofOB-cadherin primer sets include, but are not limited to: Forward5′-ACCAGATGTCTGTATCAGA-3′ (SEQ ID NO:315) and Reverse 5′-GTCTCCTGGTCATCATCTGCA-3′ (SEQ ID NO:316; Munro and Blaschuk, Biol. Reprod.55:822-827, 1996); or Forward 5′-GCCAGACACAGTTCTTAAGG-3′ (SEQ ID NO:317)and Reverse 5′-ATCAAACCTGAGTATCAGTA-3′ (SEQ ID NO:318; Goomer et al.,Calcif Tissue Int. 62:532-537, 1998). Once primer and template haveannealed, a DNA polymerase is employed to extend from the primer, thussynthesizing a copy of the template. The DNA strands are then denaturedand the process is repeated numerous times until sufficient DNA isgenerated to allow visualization by ethidium bromide staining andagarose gel electrophoresis.

[0086] Amplification may be performed on samples obtained frombiological samples taken from a test patient and an individual who isnot afflicted with a metastatic cancer. The amplification reaction maybe performed on several dilutions of cDNA spanning two orders ofmagnitude. A statistically significant (preferably at least two-fold)increase in expression in several dilutions of the test patient sampleas compared to the same dilutions of the non-cancerous sample istypically considered positive for the presence of metastatic cancer.Polynucleotide probes may also be used within in vivo diagnostic assaysperformed directly on a tumor.

[0087] Diagnostic Kits

[0088] The present invention further provides kits for use within any ofthe above diagnostic methods. Such kits typically comprise two or morecomponents necessary for performing a diagnostic assay. Components maybe compounds, reagents, containers and/or equipment. For example, onecontainer within a kit may contain a binding agent as described herein.Such binding agents may be provided attached to a support material, asdescribed above. One or more additional containers may enclose elements,such as reagents or buffers, to be used in the assay. Such kits mayalso, or alternatively, contain a detection reagent as described abovethat contains a reporter group suitable for direct or indirect detectionof binding.

[0089] Alternatively, a kit may be designed to detect the level of mRNAencoding OB-cadherin or N-cadherin in a biological sample. Such kitsgenerally comprise at least one oligonucleotide probe or primer, asdescribed above, that hybridizes to a polynucleotide encodingOB-cadherin or N-cadherin. Such an oligonucleotide may be used, forexample, within a PCR or hybridization assay. Additional components thatmay be present within such kits include a second oligonucleotide and/ora diagnostic reagent or container to facilitate the detection of apolynucleotide encoding OB-cadherin or N-cadherin.

[0090] The following Examples are offered by way of illustration and notby way of limitation.

EXAMPLES Example 1 Disruption of Human Breast Cancer Cell Adhesion

[0091] This Example illustrates the ability of a representative linearpeptide comprising an OB-cadherin cell adhesion recognition sequence todisrupt human breast epithelial cell adhesion.

[0092] MDA-MB-231 human metastatic breast cancer cells (Lombardi CancerResearch Center, Washington, D.C.) were used in these experiments. Theyexpress cadherin-11 (also known as OB-cadherin) but not N-cadherin orE-cadherin. The cells were plated (˜50,000 cells) on glass coverslipsand cultured for 24 hours in DMEM containing 5% serum. Peptides(N-Ac-IFVIDDKSG-NH₂ (SEQ ID NO:9) and H-IFVIDDKSG-OH (SEQ ID NO:3)) weredissolved in sterile water (10 mg/ml), and 100 μl of each peptide stocksolution was added to 1 ml of DMEM containing 5% serum. Control cellshad 100 μl of water added to the medium. Cells were monitored by phasecontrast microscopy. After 24 hours cells were fixed in formaldehyde.After 24 hours, neither the peptide H-IFVIDDKSG-OH (SEQ ID NO:3) norwater had an effect on cell morphology (FIG. 4A). The cells treated witheither water or H-IFVIDDKSG-OH (SEQ ID NO:3) remained flattened andwell-attached to the substratum. In contrast, the cells treated withN-Ac-IFVIDDKSG-NH₂ (SEQ ID NO:9) rounded up from each other and were notwell-attached to the substratum (FIGS. 4A and 4B; arrows indicaterounded cells). These results demonstrate that the peptideN-Ac-IFVIDDKSG-NH₂ (SEQ ID NO:9) interferes with cell adhesion. Theamino acid sequence of this peptide is identical to that which is foundin the first extracellular domain of OB-cadherin. These results indicatethat these metastatic breast cancer cells express OB-cadherin.

Example 2 Detection of OB-Cadherin in Metastatic Ovarian Tumor Cells

[0093] This Example illustrates the association between OB-cadherinexpression and metastasis in ovarian carcinoma cells.

[0094] An RT-PCR approach was employed to assay the presence ofOB-cadherin mRNA transcripts in two ovarian cancer cell lines: SKOV3 (ametastatic cell line) and OVCAR3 (a noninvasive cell line). The cDNA wassynthesized from 1 μg of total RNA by M-MLV-Reverse Transcriptase(Gibco/BRL, Burlington, ON) using a random hexamer as a primer. PCR wasperformed using the contents of the first-strand reaction and the OBcadherin-specific primers and Taq polymerase (Boehringer Mannheim,Laval, Que., Canada). The OB-cadherin-specific primers used were:Forward 5′-ACCAGATGTCTGTATCAGA3′; (SEQ ID NO:315) and Reverse5′-GTCTCCTGGTCATCATCTGCA-3′ (SEQ ID NO:316)

[0095] (Munro and Blaschuk, Biol. Reprod. 55:822-827, 1996). To confirmthe quality of the RNA used, PCR was also performed using primers forthe housekeeping gene, hypoxanthine phosphoribosyltransferase (HPRT).The HPRT-specific primers used were: Forward5′-CCTGCTGGATTACATTAAAGCACTG-3′; (SEQ ID NO:319) and Reverse5′-GTCAAGGGCATATCCAACAACAAAC-3′ (SEQ ID NO:320)

[0096] (Melton et al., Proc. Natl. Acad. Sci. USA 81:2147-2151, 1984).The cycling program was as follows: denaturation at 95° C. for 30 sec.;annealing at 58-60° C. for 45 sec.; polymerization at 72° C. for 1 min.;repeat for 30 cycles. All PCR reactions were performed in parallel withreactions containing no cDNA as a control for contamination of PCRreagents. Products were identified by agarose gel electrophoresisstained with ethidium bromide (Sambrook et al., Molecular Cloning: ALaboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor,N.Y., 1989).

[0097] The results are presented in FIG. 5, which shows RT-PCR productsfrom SKOV3 (lane 1) and OVCAR3 (lane 2). The primers used are specificfor OB-cadherin (OB-cad) and hypoxanthine phosphoribosyltransferase(HPRT) as indicated, with an expected PCR product of 745 bp and 352 bp,respectively. Products were stained with ethidium bromide and resolvedby agarose gel electrophoresis, and were all of the expected size. Theresults indicate that OB-cadherin is expressed by metastatic ovariancancer cells, and is not expressed by non-invasive ovarian cancer cells.

Example 3 Detection of OB-Cadherin in Leukemic Cells

[0098] This Example illustrates the expression of OB-cadherin inlymphocytes of leukemia patients.

[0099] The RT-PCR approach described in Example 2 was employed to assaythe presence of OB-cadherin mRNA transcripts in lymphocytes extractedfrom patients with B-cell chronic lymphocytic leukemia (B-CLL). RT-PCRproducts (shown in FIG. 6) were generated from lymphocytes of a humanB-CLL patient (lane 1) and mouse liver (lane 2). The primers used werespecific for OB-cadherin (OB-cad, top panel) and hypoxanthinephosphoribosyltransferase (HPRT, bottom panel), with an expected PCRproduct of 745 bp and 352 bp, respectively. Products were stained withethidium bromide and resolved by agarose gel electrophoresis, and wereall of the expected size. The results indicate that lymphocytes of aleukemia patient express OB-cadherin.

Example 4 Expression of N-Cadherin in Metastatic Carcinoma Cells

[0100] This Example illustrates the correlation between N-cadherin andmetastatic potential in ovarian carcinoma cell lines.

[0101] E-cadherin and N-cadherin expression was evaluated in a series ofovarian carcinoma cell lines, using the RT-PCR approach described above.The E-cadherin specific primer used were: Forward5′-CCTTCCCCCAACACGTCCCCCC-3′; (SEQ ID NO:321) and Reverse5′-TCTCCACCTCCTTCTTCATC-3′ (SEQ ID NO:322)

[0102] (Munro and Blaschuk, Biol. Reprod 55:822-827, 1996). TheN-cadherin specific primers used were: Forward5′-CAAGAGCTTGTCACAATCAGG-3′; (SEQ ID NO:323) and Reverse5′-CATTTGGATCATCCGCATC-3′ (SEQ ID NO:324)

[0103] (Munro and Blaschuk, Biol. Reprod. 55:822-827, 1996).

[0104] Cell lines examined included OVCAR-3 (Hamilton et al., CancerResearch 43:5379-89,1983); SW626 (Ripamonti et al., Cancer Immunology,Immunotherapy 24:13-18, 1987); CaOV3, SKOV3 and HEY (Buick et al.,Cancer Research 45:3668-76, 1985). These cells (except HEY) are alsoavailable from American Type Culture Collection (Manassas, Va.).

[0105] The results of these analyses are presented in Table I, below, inwhich detectable PCR product is indicated as a “+” and no detectable PCRproduct is indicated by a “−”. TABLE 1 N- and E-Cadherin Expression inOvarian Carcinoma Cell Lines Differentiation Stage Cadherin Cell LinePhenotype and Metastatic Potential E N Normal Epithelial None + −OVCAR-3 Adenocarcinoma Differentiated; low + − metastatic SW626Adenocarcinoma Differentiated; low + − metastatic CaOV3 Adenocarcinoma? + + SKOV3 Adenocarcinoma Poor differentaiation; − + high metastaticHEY Adenocarcinoma Poor differentiation; − + high metastatic

[0106] From the foregoing it will be appreciated that, although specificembodiments of the invention have been described herein for purposes ofillustration, various modifications may be made without deviating fromthe spirit and scope of the invention. Accordingly, the invention is notlimited except as by the appended claims.

1 324 1 5 PRT Unknown MOD_RES (2) Where Xaa is any amino acid 1 Asp XaaAsn Asp Asn 1 5 2 4 PRT Unknown Description of Unknown Organism CalciumBinding Motif in Extracellular domains of Classical Cadherins 2 Leu AspArg Glu 1 3 9 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis based on Human OB-Cadherin 3 Ile Phe Val Ile AspAsp Lys Ser Gly 1 5 4 106 PRT Homo sapiens 4 Gly Trp Val Trp Asn Gln PhePhe Val Ile Glu Glu Tyr Thr Gly Pro 1 5 10 15 Asp Pro Val Leu Val GlyArg Leu His Ser Asp Ile Asp Ser Gly Asp 20 25 30 Gly Asn Ile Lys Tyr IleLeu Ser Gly Glu Gly Ala Gly Thr Ile Phe 35 40 45 Val Ile Asp Asp Lys SerGly Asn Ile His Ala Thr Lys Thr Leu Asp 50 55 60 Arg Glu Glu Arg Ala GlnTyr Thr Leu Met Ala Gln Ala Val Asp Arg 65 70 75 80 Asp Thr Asn Arg ProLeu Glu Pro Pro Ser Glu Phe Ile Val Lys Val 85 90 95 Gln Asp Ile Asn AspAsn Pro Pro Glu Phe 100 105 5 106 PRT Mus musculus 5 Gly Trp Val Trp AsnGln Phe Phe Val Ile Glu Glu Tyr Thr Gly Pro 1 5 10 15 Asp Pro Val LeuVal Gly Arg Leu His Ser Asp Ile Asp Ser Gly Asp 20 25 30 Gly Asn Ile LysTyr Ile Leu Ser Gly Glu Gly Ala Gly Thr Ile Phe 35 40 45 Val Ile Asp AspLys Ser Gly Asn Ile His Ala Thr Lys Thr Leu Asp 50 55 60 Arg Glu Glu ArgAla Gln Tyr Thr Leu Met Ala Gln Ala Val Asp Arg 65 70 75 80 Asp Thr AsnArg Pro Leu Glu Pro Pro Ser Glu Phe Ile Val Lys Val 85 90 95 Gln Asp IleAsn Asp Asn Pro Pro Glu Phe 100 105 6 108 PRT Homo sapiens 6 Asp Trp ValIle Pro Pro Ile Asn Leu Pro Glu Asn Ser Arg Gly Pro 1 5 10 15 Phe ProGln Glu Leu Val Arg Ile Arg Ser Asp Arg Asp Lys Asn Leu 20 25 30 Ser LeuArg Tyr Ser Val Thr Gly Pro Gly Ala Asp Gln Pro Pro Thr 35 40 45 Gly IlePhe Ile Leu Asn Pro Ile Ser Gly Gln Leu Ser Val Thr Lys 50 55 60 Pro LeuAsp Arg Glu Gln Ile Ala Arg Phe His Leu Arg Ala His Ala 65 70 75 80 ValAsp Ile Asn Gly Asn Gln Val Glu Asn Pro Ile Asp Ile Val Ile 85 90 95 AsnVal Ile Asp Met Asn Asp Asn Arg Pro Glu Phe 100 105 7 108 PRT Musmusculus 7 Asp Trp Val Ile Pro Pro Ile Asn Leu Pro Glu Asn Ser Arg GlyPro 1 5 10 15 Phe Pro Gln Glu Leu Val Arg Ile Arg Ser Asp Arg Asp LysAsn Leu 20 25 30 Ser Leu Arg Tyr Ser Val Thr Gly Pro Gly Ala Asp Gln ProPro Thr 35 40 45 Gly Ile Phe Ile Ile Asn Pro Ile Ser Gly Gln Leu Ser ValThr Lys 50 55 60 Pro Leu Asp Arg Glu Leu Ile Ala Arg Phe His Leu Arg AlaHis Ala 65 70 75 80 Val Asp Ile Asn Gly Asn Gln Val Glu Asn Pro Ile AspIle Val Ile 85 90 95 Asn Val Ile Asp Met Asn Asp Asn Arg Pro Glu Phe 100105 8 108 PRT Bos taurus 8 Asp Trp Val Ile Pro Pro Ile Asn Leu Pro GluAsn Ser Arg Gly Pro 1 5 10 15 Phe Pro Gln Glu Leu Val Arg Ile Arg SerAsp Arg Asp Lys Asn Leu 20 25 30 Ser Leu Arg Tyr Ser Val Thr Gly Pro GlyAla Asp Gln Pro Pro Thr 35 40 45 Gly Ile Phe Ile Ile Asn Pro Ile Ser GlyGln Leu Ser Val Thr Lys 50 55 60 Pro Leu Asp Arg Glu Leu Ile Ala Arg PheHis Leu Arg Ala His Ala 65 70 75 80 Val Asp Ile Asn Gly Asn Gln Val GluAsn Pro Ile Asp Ile Val Ile 85 90 95 Asn Val Ile Asp Met Asn Asp Asn ArgPro Glu Phe 100 105 9 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis based on Human OB-Cadherin 9Ile Phe Val Ile Asp Asp Lys Ser Gly 1 5 10 9 PRT Unknown Description ofUnknown Organism Consensus Cell Adhesion Recognition Sequence in anOB-Cadherin 10 Xaa Phe Xaa Xaa Xaa Xaa Xaa Xaa Gly 1 5 11 4 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 11 Ile Asp Asp Lys 1 12 4 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 12 Asp Asp Lys Ser 1 13 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 13 Val Ile Asp Asp Lys 1 5 14 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 14 Ile Asp Asp Lys Ser 1 5 15 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 15 Val Ile Asp Asp Lys Ser 1 5 16 5PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 16 Asp Asp Lys Ser Gly 1 5 17 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 17 Ile Asp Asp Lys Ser Gly 1 5 18 7PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 18 Val Ile Asp Asp Lys Ser Gly 1 519 6 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis based on Human OB-Cadherin 19 Phe Val Ile Asp Asp Lys 1 520 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis based on Human OB-Cadherin 20 Phe Val Ile Asp Asp Lys Ser 15 21 8 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis based on Human OB-Cadherin 21 Phe Val Ile Asp AspLys Ser Gly 1 5 22 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis based on Human OB-Cadherin 22 Ile Phe ValIle Asp Asp Lys 1 5 23 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis based on Human OB-Cadherin 23Ile Phe Val Ile Asp Asp Lys Ser 1 5 24 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis based on HumanOB-Cadherin 24 Ile Phe Val Ile Asp Asp Lys Ser Gly 1 5 25 4 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 25 Ile Glu Glu Tyr 1 26 4 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 26 Glu Glu Tyr Thr 1 27 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 27 Val Ile Glu Glu Tyr 1 5 28 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 28 Ile Glu Glu Tyr Thr 1 5 29 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 29 Val Ile Glu Glu Tyr Thr 1 5 30 5PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 30 Glu Glu Tyr Thr Gly 1 5 31 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 31 Ile Glu Glu Tyr Thr Gly 1 5 32 7PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 32 Val Ile Glu Glu Tyr Thr Gly 1 533 6 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis based on Human OB-Cadherin 33 Phe Val Ile Glu Glu Tyr 1 534 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis based on Human OB-Cadherin 34 Phe Val Ile Glu Glu Tyr Thr 15 35 8 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis based on Human OB-Cadherin 35 Phe Val Ile Glu GluTyr Thr Gly 1 5 36 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis based on Human OB-Cadherin 36 Phe Phe ValIle Glu Glu Tyr 1 5 37 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis based on Human OB-Cadherin 37Phe Phe Val Ile Glu Glu Tyr Thr 1 5 38 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis based on HumanOB-Cadherin 38 Phe Phe Val Ile Glu Glu Tyr Thr Gly 1 5 39 4 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 39 Val Glu Ala Gln 1 40 4 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 40 Glu Ala Gln Thr 1 41 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 41 Ser Val Glu Ala Gln 1 5 42 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 42 Val Glu Ala Gln Thr 1 5 43 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 43 Ser Val Glu Ala Gln Thr 1 5 44 5PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 44 Glu Ala Gln Thr Gly 1 5 45 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 45 Val Glu Ala Gln Thr Gly 1 5 46 7PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 46 Ser Val Glu Ala Gln Thr Gly 1 547 6 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis based on Human OB-Cadherin 47 Phe Ser Val Glu Ala Gln 1 548 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis based on Human OB-Cadherin 48 Phe Ser Val Glu Ala Gln Thr 15 49 8 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis based on Human OB-Cadherin 49 Phe Ser Val Glu AlaGln Thr Gly 1 5 50 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis based on Human OB-Cadherin 50 Tyr Phe SerVal Glu Ala Gln 1 5 51 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis based on Human OB-Cadherin 51Tyr Phe Ser Val Glu Ala Gln Thr 1 5 52 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis based on HumanOB-Cadherin 52 Tyr Phe Ser Val Glu Ala Gln Thr Gly 1 5 53 9 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis based on Human OB-Cadherin 53 Phe Phe Val Ile Glu Glu Tyr ThrGly 1 5 54 9 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis based on Human OB-Cadherin 54 Tyr Phe Ser Val GluAla Gln Thr Gly 1 5 55 5 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 55 Cys Asp Asp Lys Cys 1 5 56 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 56 Cys Ile Asp Asp Lys Cys 1 5 57 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 57 Cys Asp Asp LysSer Cys 1 5 58 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin58 Cys Val Ile Asp Asp Lys Cys 1 5 59 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 59 Cys Ile Asp Asp Lys Ser Cys 1 5 60 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 60 Cys Val Ile AspAsp Lys Ser Cys 1 5 61 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 61 Cys Asp Asp Lys Ser Gly Cys 1 5 62 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 62 Cys Ile Asp Asp Lys Ser GlyCys 1 5 63 9 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 63 CysVal Ile Asp Asp Lys Ser Gly Cys 1 5 64 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 64 Cys Phe Val Ile Asp Asp Lys Cys 1 5 65 9PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 65 Cys Phe Val IleAsp Asp Lys Ser Cys 1 5 66 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 66 Cys Phe Val Ile Asp Asp Lys Ser Gly Cys 1 5 10 67 9 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 67 Cys Ile Phe ValIle Asp Asp Lys Cys 1 5 68 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 68 Cys Ile Phe Val Ile Asp Asp Lys Ser Cys 1 5 10 69 11 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 69 Cys Ile Phe ValIle Asp Asp Lys Ser Gly Cys 1 5 10 70 5 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 70 Asp Asp Asp Lys Lys 1 5 71 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 71 Asp Ile Asp AspLys Lys 1 5 72 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin72 Asp Val Ile Asp Asp Lys Lys 1 5 73 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 73 Asp Phe Val Ile Asp Asp Lys Lys 1 5 74 9PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 74 Asp Ile Phe ValIle Asp Asp Lys Lys 1 5 75 5 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 75 Glu Asp Asp Lys Lys 1 5 76 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 76 Glu Ile Asp Asp Lys Lys 1 5 77 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 77 Glu Val Ile AspAsp Lys Lys 1 5 78 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin78 Glu Phe Val Ile Asp Asp Lys Lys 1 5 79 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 79 Glu Ile Phe Val Ile Asp Asp Lys Lys 1 5 806 PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 80 Phe Val Ile AspAsp Lys 1 5 81 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin81 Phe Val Ile Asp Asp Lys Ser 1 5 82 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 82 Phe Val Ile Asp Asp Lys Ser Gly 1 5 83 5PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 83 Lys Asp Asp LysAsp 1 5 84 6 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 84 LysIle Asp Asp Lys Asp 1 5 85 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 85 Lys Asp Asp Lys Ser Asp 1 5 86 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 86 Lys Val Ile Asp Asp Lys Asp 1 5 87 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 87 Lys Ile Asp AspLys Ser Asp 1 5 88 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin88 Lys Val Ile Asp Asp Lys Ser Asp 1 5 89 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 89 Lys Asp Asp Lys Ser Gly Asp 1 5 90 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 90 Lys Ile Asp AspLys Ser Gly Asp 1 5 91 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 91 Lys Val Ile Asp Asp Lys Ser Gly Asp 1 5 92 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 92 Lys Phe Val IleAsp Asp Lys Asp 1 5 93 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 93 Lys Phe Val Ile Asp Asp Lys Ser Asp 1 5 94 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 94 Lys Phe Val IleAsp Asp Lys Ser Gly Asp 1 5 10 95 9 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 95 Lys Ile Phe Val Ile Asp Asp Lys Asp 1 5 96 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 96 Lys Ile Phe ValIle Asp Asp Lys Ser Asp 1 5 10 97 11 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 97 Lys Ile Phe Val Ile Asp Asp Lys Ser Gly Asp 1 5 1098 5 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 98 Val Ile AspAsp Lys 1 5 99 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin99 Ile Asp Asp Lys Ser 1 5 100 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 100 Val Ile Asp Asp Lys Ser 1 5 101 7 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 101 Val Ile Asp Asp Lys Ser Gly 15 102 5 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 102 AspAsp Lys Ser Gly 1 5 103 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 103 Ile Asp Asp Lys Ser Gly 1 5 104 7 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 104 Ile Phe Val Ile Asp Asp Lys 15 105 8 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 105 IlePhe Val Ile Asp Asp Lys Ser 1 5 106 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 106 Ile Phe Val Ile Asp Asp Lys Ser Gly 1 5107 5 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 107 Lys Asp AspLys Glu 1 5 108 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin108 Lys Ile Asp Asp Lys Glu 1 5 109 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 109 Lys Asp Asp Lys Ser Glu 1 5 110 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 110 Lys Val Ile AspAsp Lys Glu 1 5 111 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin111 Lys Ile Asp Asp Lys Ser Glu 1 5 112 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 112 Lys Val Ile Asp Asp Lys Ser Glu 1 5 113 7PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 113 Lys Asp Asp LysSer Gly Glu 1 5 114 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin114 Lys Ile Asp Asp Lys Ser Gly Glu 1 5 115 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 115 Lys Val Ile Asp Asp Lys Ser Gly Glu 1 5116 8 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 116 Lys Phe ValIle Asp Asp Lys Glu 1 5 117 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 117 Lys Phe Val Ile Asp Asp Lys Ser Glu 1 5 118 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 118 Lys Phe Val IleAsp Asp Lys Ser Gly Glu 1 5 10 119 9 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 119 Lys Ile Phe Val Ile Asp Asp Lys Glu 1 5 120 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 120 Lys Ile Phe ValIle Asp Asp Lys Ser Glu 1 5 10 121 11 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 121 Lys Ile Phe Val Ile Asp Asp Lys Ser GlyGlu 1 5 10 122 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin122 Cys Glu Glu Tyr Cys 1 5 123 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 123 Cys Ile Glu Glu Tyr Cys 1 5 124 6 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 124 Cys Glu Glu Tyr Thr Cys 1 5125 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 125 Cys Val IleGlu Glu Tyr Cys 1 5 126 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 126 Cys Ile Glu Glu Tyr Thr Cys 1 5 127 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 127 Cys Val Ile Glu Glu Tyr ThrCys 1 5 128 7 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 128 CysGlu Glu Tyr Thr Gly Cys 1 5 129 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 129 Cys Ile Glu Glu Tyr Thr Gly Cys 1 5 130 9 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 130 Cys Val Ile Glu Glu Tyr ThrGly Cys 1 5 131 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin131 Cys Phe Val Ile Glu Glu Tyr Cys 1 5 132 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 132 Cys Phe Val Ile Glu Glu Tyr Thr Cys 1 5133 10 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 133 CysPhe Val Ile Glu Glu Tyr Thr Gly Cys 1 5 10 134 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 134 Cys Phe Phe Val Ile Glu Glu Tyr Cys 1 5135 10 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 135 CysPhe Phe Val Ile Glu Glu Tyr Thr Cys 1 5 10 136 11 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 136 Cys Phe Phe Val Ile Glu GluTyr Thr Gly Cys 1 5 10 137 5 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 137 Lys Glu Glu Tyr Asp 1 5 138 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 138 Lys Ile Glu Glu Tyr Asp 1 5 139 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 139 Lys Glu Glu TyrThr Asp 1 5 140 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin140 Lys Val Ile Glu Glu Tyr Asp 1 5 141 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 141 Lys Ile Glu Glu Tyr Thr Asp 1 5 142 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 142 Lys Val Ile GluGlu Tyr Thr Asp 1 5 143 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 143 Lys Glu Glu Tyr Thr Gly Cys Asp 1 5 144 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 144 Lys Ile Glu Glu Tyr Thr GlyAsp 1 5 145 9 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 145 LysVal Ile Glu Glu Tyr Thr Gly Asp 1 5 146 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 146 Lys Phe Val Ile Glu Glu Tyr Asp 1 5 147 9PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 147 Lys Phe Val IleGlu Glu Tyr Thr Asp 1 5 148 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 148 Lys Phe Val Ile Glu Glu Tyr Thr Gly Asp 1 5 10 149 9 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 149 Lys Phe Phe ValIle Glu Glu Tyr Asp 1 5 150 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 150 Lys Phe Phe Val Ile Glu Glu Tyr Thr Asp 1 5 10 151 11PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 151 Lys Phe Phe ValIle Glu Glu Tyr Thr Gly Asp 1 5 10 152 5 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 152 Glu Glu Glu Tyr Lys 1 5 153 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 153 Glu Ile Glu GluTyr Lys 1 5 154 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin154 Glu Glu Glu Tyr Thr Lys 1 5 155 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 155 Glu Val Ile Glu Glu Tyr Lys 1 5 156 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 156 Glu Ile Glu GluTyr Thr Lys 1 5 157 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin157 Glu Val Ile Glu Glu Tyr Thr Lys 1 5 158 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 158 Glu Glu Glu Tyr Thr Gly Lys 1 5 159 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 159 Glu Ile Glu GluTyr Thr Gly Lys 1 5 160 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 160 Glu Val Ile Glu Glu Tyr Thr Gly Lys 1 5 161 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 161 Glu Phe Val IleGlu Glu Tyr Lys 1 5 162 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 162 Glu Phe Val Ile Glu Glu Tyr Thr Lys 1 5 163 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 163 Glu Phe Val IleGlu Glu Tyr Thr Gly Lys 1 5 10 164 9 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 164 Glu Phe Phe Val Ile Glu Glu Tyr Lys 1 5 165 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 165 Glu Phe Phe ValIle Glu Glu Tyr Thr Lys 1 5 10 166 11 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 166 Glu Phe Phe Val Ile Glu Glu Tyr Thr GlyLys 1 5 10 167 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin167 Asp Cys Glu Glu Tyr Lys 1 5 168 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 168 Asp Ile Glu Glu Tyr Cys Lys 1 5 169 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 169 Asp Glu Glu TyrThr Lys 1 5 170 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin170 Asp Val Ile Glu Glu Tyr Lys 1 5 171 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 171 Asp Ile Glu Glu Tyr Thr Lys 1 5 172 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 172 Asp Val Ile GluGlu Tyr Thr Lys 1 5 173 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 173 Asp Glu Glu Tyr Thr Gly Lys 1 5 174 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 174 Asp Ile Glu Glu Tyr Thr GlyLys 1 5 175 9 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 175 AspVal Ile Glu Glu Tyr Thr Gly Lys 1 5 176 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 176 Asp Phe Val Ile Glu Glu Tyr Lys 1 5 177 9PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 177 Asp Phe Val IleGlu Glu Tyr Thr Lys 1 5 178 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 178 Asp Phe Val Ile Glu Glu Tyr Thr Gly Lys 1 5 10 179 9 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 179 Asp Phe Phe ValIle Glu Glu Tyr Lys 1 5 180 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 180 Asp Phe Phe Val Ile Glu Glu Tyr Thr Lys 1 5 10 181 11PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 181 Asp Phe Phe ValIle Glu Glu Tyr Thr Gly Lys 1 5 10 182 5 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 182 Lys Glu Glu Tyr Glu 1 5 183 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 183 Lys Ile Glu GluTyr Glu 1 5 184 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin184 Lys Glu Glu Tyr Thr Glu 1 5 185 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 185 Lys Val Ile Glu Glu Tyr Glu 1 5 186 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 186 Lys Ile Glu GluTyr Thr Glu 1 5 187 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin187 Lys Val Ile Glu Glu Tyr Thr Glu 1 5 188 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 188 Lys Glu Glu Tyr Thr Gly Glu 1 5 189 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 189 Lys Ile Glu GluTyr Thr Gly Glu 1 5 190 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 190 Lys Val Ile Glu Glu Tyr Thr Gly Glu 1 5 191 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 191 Lys Phe Val IleGlu Glu Tyr Glu 1 5 192 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 192 Lys Phe Val Ile Glu Glu Tyr Thr Glu 1 5 193 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 193 Lys Phe Val IleGlu Glu Tyr Thr Gly Glu 1 5 10 194 9 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 194 Lys Phe Phe Val Ile Glu Glu Tyr Glu 1 5 195 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 195 Lys Phe Phe ValIle Glu Glu Tyr Thr Glu 1 5 10 196 11 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 196 Lys Phe Phe Val Ile Glu Glu Tyr Thr GlyGlu 1 5 10 197 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin197 Val Ile Glu Glu Tyr 1 5 198 5 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 198 Ile Glu Glu Tyr Thr 1 5 199 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 199 Val Ile Glu Glu Tyr Thr 1 5 200 5 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 200 Glu Glu Tyr ThrGly 1 5 201 6 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 201 IleGlu Glu Tyr Thr Gly 1 5 202 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 202 Val Ile Glu Glu Tyr Thr Gly 1 5 203 6 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 203 Phe Val Ile Glu Glu Tyr 1 5204 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 204 Phe Val IleGlu Glu Tyr Thr 1 5 205 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 205 Phe Val Ile Glu Glu Tyr Thr Gly 1 5 206 7 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 206 Phe Phe Val Ile Glu Glu Tyr 15 207 8 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 207 PhePhe Val Ile Glu Glu Tyr Thr 1 5 208 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 208 Phe Phe Val Ile Glu Glu Tyr Thr Gly 1 5209 5 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 209 Cys Glu AlaGln Cys 1 5 210 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin210 Cys Val Glu Ala Gln Cys 1 5 211 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 211 Cys Glu Ala Gln Thr Cys 1 5 212 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 212 Cys Ser Val GluAla Gln Cys 1 5 213 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin213 Cys Val Glu Ala Gln Thr Cys 1 5 214 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 214 Cys Ser Val Glu Ala Gln Thr Cys 1 5 215 7PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 215 Cys Glu Ala GlnThr Gly Cys 1 5 216 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin216 Cys Val Glu Ala Gln Thr Gly Cys 1 5 217 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 217 Cys Ser Val Glu Ala Gln Thr Gly Cys 1 5218 8 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 218 Cys Phe SerVal Glu Ala Gln Cys 1 5 219 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 219 Cys Phe Ser Val Glu Ala Gln Thr Cys 1 5 220 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 220 Cys Phe Ser ValGlu Ala Gln Thr Gly Cys 1 5 10 221 9 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 221 Cys Tyr Phe Ser Val Glu Ala Gln Cys 1 5 222 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 222 Cys Tyr Phe SerVal Glu Ala Gln Thr Cys 1 5 10 223 11 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 223 Cys Tyr Phe Ser Val Glu Ala Gln Thr GlyCys 1 5 10 224 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin224 Lys Glu Ala Gln Asp 1 5 225 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 225 Lys Val Glu Ala Gln Asp 1 5 226 6 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 226 Lys Glu Ala Gln Thr Asp 1 5227 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 227 Lys Ser ValGlu Ala Gln Asp 1 5 228 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 228 Lys Val Glu Ala Gln Thr Asp 1 5 229 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 229 Lys Ser Val Glu Ala Gln ThrAsp 1 5 230 7 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 230 LysGlu Ala Gln Thr Gly Asp 1 5 231 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 231 Lys Val Glu Ala Gln Thr Gly Asp 1 5 232 9 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 232 Lys Ser Val Glu Ala Gln ThrGly Asp 1 5 233 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin233 Lys Phe Ser Val Glu Ala Gln Asp 1 5 234 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 234 Lys Phe Ser Val Glu Ala Gln Thr Asp 1 5235 10 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 235 LysPhe Ser Val Glu Ala Gln Thr Gly Asp 1 5 10 236 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 236 Lys Tyr Phe Ser Val Glu Ala Gln Asp 1 5237 10 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 237 LysTyr Phe Ser Val Glu Ala Gln Thr Asp 1 5 10 238 11 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 238 Lys Tyr Phe Ser Val Glu AlaGln Thr Gly Asp 1 5 10 239 5 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 239 Glu Glu Ala Gln Lys 1 5 240 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 240 Glu Val Glu Ala Gln Lys 1 5 241 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 241 Glu Glu Ala GlnThr Lys 1 5 242 7 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin242 Glu Ser Val Glu Ala Gln Lys 1 5 243 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 243 Glu Val Glu Ala Gln Thr Lys 1 5 244 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 244 Glu Ser Val GluAla Gln Thr Lys 1 5 245 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 245 Glu Glu Ala Gln Thr Gly Lys 1 5 246 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 246 Glu Val Glu Ala Gln Thr GlyLys 1 5 247 9 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 247 GluSer Val Glu Ala Gln Thr Gly Lys 1 5 248 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 248 Glu Phe Ser Val Glu Ala Gln Lys 1 5 249 9PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 249 Glu Phe Ser ValGlu Ala Gln Thr Lys 1 5 250 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 250 Glu Phe Ser Val Glu Ala Gln Thr Gly Lys 1 5 10 251 9 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 251 Glu Tyr Phe SerVal Glu Ala Gln Lys 1 5 252 10 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 252 Glu Tyr Phe Ser Val Glu Ala Gln Thr Lys 1 5 10 253 11PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 253 Glu Tyr Phe SerVal Glu Ala Gln Thr Gly Lys 1 5 10 254 5 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 254 Asp Glu Ala Gln Lys 1 5 255 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 255 Asp Val Glu AlaGln Lys 1 5 256 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin256 Asp Glu Ala Gln Thr Lys 1 5 257 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 257 Asp Ser Val Glu Ala Gln Lys 1 5 258 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 258 Asp Val Glu AlaGln Thr Lys 1 5 259 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin259 Asp Ser Val Glu Ala Gln Thr Lys 1 5 260 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 260 Asp Glu Ala Gln Thr Gly Lys 1 5 261 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 261 Asp Val Glu AlaGln Thr Gly Lys 1 5 262 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 262 Asp Ser Val Glu Ala Gln Thr Gly Lys 1 5 263 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 263 Asp Phe Ser ValGlu Ala Gln Lys 1 5 264 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 264 Asp Phe Ser Val Glu Ala Gln Thr Lys 1 5 265 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 265 Asp Phe Ser ValGlu Ala Gln Thr Gly Lys 1 5 10 266 9 PRT Artificial Sequence Descriptionof Artificial Sequence Product of Synthesis and Cyclization based onHuman OB-Cadherin 266 Asp Tyr Phe Ser Val Glu Ala Gln Lys 1 5 267 10 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 267 Asp Tyr Phe SerVal Glu Ala Gln Thr Lys 1 5 10 268 11 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 268 Asp Tyr Phe Ser Val Glu Ala Gln Thr GlyLys 1 5 10 269 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin269 Lys Glu Ala Gln Glu 1 5 270 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 270 Lys Val Glu Ala Gln Glu 1 5 271 6 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 271 Lys Glu Ala Gln Thr Glu 1 5272 7 PRT Artificial Sequence Description of Artificial Sequence Productof Synthesis and Cyclization based on Human OB-Cadherin 272 Lys Ser ValGlu Ala Gln Glu 1 5 273 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 273 Lys Val Glu Ala Gln Thr Glu 1 5 274 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 274 Lys Ser Val Glu Ala Gln ThrGlu 1 5 275 7 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 275 LysGlu Ala Gln Thr Gly Glu 1 5 276 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 276 Lys Val Glu Ala Gln Thr Gly Glu 1 5 277 9 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 277 Lys Ser Val Glu Ala Gln ThrGly Glu 1 5 278 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin278 Lys Phe Ser Val Glu Ala Gln Glu 1 5 279 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 279 Lys Phe Ser Val Glu Ala Gln Thr Glu 1 5280 10 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 280 LysPhe Ser Val Glu Ala Gln Thr Gly Glu 1 5 10 281 9 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 281 Lys Tyr Phe Ser Val Glu Ala Gln Glu 1 5282 10 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 282 LysTyr Phe Ser Val Glu Ala Gln Thr Glu 1 5 10 283 11 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 283 Lys Tyr Phe Ser Val Glu AlaGln Thr Gly Glu 1 5 10 284 5 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 284 Ser Val Glu Ala Gln 1 5 285 5 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human OB-Cadherin 285 Val Glu Ala Gln Thr 1 5 286 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human OB-Cadherin 286 Ser Val Glu AlaGln Thr 1 5 287 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human OB-Cadherin287 Glu Ala Gln Thr Gly 1 5 288 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 288 Val Glu Ala Gln Thr Gly 1 5 289 7 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 289 Ser Val Glu Ala Gln Thr Gly 15 290 6 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 290 PheSer Val Glu Ala Gln 1 5 291 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 291 Phe Ser Val Glu Ala Gln Thr 1 5 292 8 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 292 Phe Ser Val Glu Ala Gln ThrGly 1 5 293 7 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human OB-Cadherin 293 TyrPhe Ser Val Glu Ala Gln 1 5 294 8 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanOB-Cadherin 294 Tyr Phe Ser Val Glu Ala Gln Thr 1 5 295 9 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human OB-Cadherin 295 Tyr Phe Ser Val Glu Ala GlnThr Gly 1 5 296 5 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human N-Cadherin296 Cys His Ala Val Cys 1 5 297 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanN-Cadherin 297 Cys His Ala Val Asp Cys 1 5 298 6 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human N-Cadherin 298 Cys Ala His Ala Val Cys 1 5 299 7 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human N-Cadherin 299 Cys Ala His AlaVal Asp Cys 1 5 300 8 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human N-Cadherin300 Cys Ala His Ala Val Asp Ile Cys 1 5 301 8 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human N-Cadherin 301 Cys Arg Ala His Ala Val Asp Cys 1 5 302 8PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human N-Cadherin 302 Cys Leu Arg AlaHis Ala Val Cys 1 5 303 9 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanN-Cadherin 303 Cys Leu Arg Ala His Ala Val Asp Cys 1 5 304 6 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human N-Cadherin 304 Cys Ser His AlaVal Cys 1 5 305 6 PRT Artificial Sequence Description of ArtificialSequence Product of Synthesis and Cyclization based on Human N-Cadherin305 Cys His Ala Val Ser Cys 1 5 306 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human N-Cadherin 306 Cys Ser His Ala Val Ser Cys 1 5 307 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human N-Cadherin 307 Cys Ser His AlaVal Ser Ser Cys 1 5 308 7 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanN-Cadherin 308 Cys His Ala Val Ser Ser Cys 1 5 309 5 PRT ArtificialSequence Description of Artificial Sequence Product of Synthesis andCyclization based on Human N-Cadherin 309 Lys His Ala Val Asp 1 5 310 5PRT Artificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human N-Cadherin 310 Asp His Ala ValLys 1 5 311 5 PRT Artificial Sequence Description of Artificial SequenceProduct of Synthesis and Cyclization based on Human N-Cadherin 311 LysHis Ala Val Glu 1 5 312 6 PRT Artificial Sequence Description ofArtificial Sequence Product of Synthesis and Cyclization based on HumanN-Cadherin 312 Ala His Ala Val Asp Ile 1 5 313 7 PRT Artificial SequenceDescription of Artificial Sequence Product of Synthesis and Cyclizationbased on Human N-Cadherin 313 Ser His Ala Val Asp Ser Ser 1 5 314 8 PRTArtificial Sequence Description of Artificial Sequence Product ofSynthesis and Cyclization based on Human N-Cadherin 314 Lys Ser His AlaVal Ser Ser Asp 1 5 315 19 DNA Artificial Sequence Description ofArtificial Sequence OB-Cadherin forward primer 315 accagatgtc tgtatcaga19 316 21 DNA Artificial Sequence Description of Artificial SequenceOB-Cadherin reverse primer 316 gtctcctggt catcatctgc a 21 317 20 DNAArtificial Sequence Description of Artificial Sequence OB-Cadherinforward primer 317 gccagacaca gttcttaagg 20 318 19 DNA ArtificialSequence Description of Artificial Sequence OB-Cadherin reverse primer318 atcaacctga gtatcagta 19 319 25 DNA Artificial Sequence Descriptionof Artificial Sequence Hypoxanthine phosphoribosyltransferase(HPRT)-specific forward primer 319 cctgctggat tacattaaag cactg 25 320 25DNA Artificial Sequence Description of Artificial Sequence Hypoxanthinephosphoribosyltransferase (HPRT)-specific reverse primer 320 gtcaagggcatatccaacaa caaac 25 321 22 DNA Artificial Sequence Description ofArtificial Sequence E-Cadherin forward primer 321 ccttccccca acacgtcccccc 22 322 20 DNA Artificial Sequence Description of Artificial SequenceE-Cadherin reverse primer 322 tctccacctc cttcttcatc 20 323 21 DNAArtificial Sequence Description of Artificial Sequence N-Cadherinforward primer 323 caagagcttg tcacaatcag g 21 324 19 DNA ArtificialSequence Description of Artificial Sequence N-Cadherin reverse primer324 catttggatc atccgcatc 19

1. A method for determining the presence or absence of a cancer in apatient, comprising the steps of: (a) contacting a biological sampleobtained from a patient with a binding agent that specifically binds toOB-cadherin; and (b) detecting in the sample an amount of polypeptidethat binds to the binding agent, relative to a predetermined cut-offvalue, and therefrom determining the presence or absence of a cancer inthe patient.
 2. A method according to claim 1 wherein the binding agentis a monoclonal antibody.
 3. A method according to claim 1 wherein thebinding agent is a polyclonal antibody.
 4. A method according to claim1, wherein the binding agent comprises an OB-cadherin CAR sequence, oran analogue or mimetic thereof.
 5. A method according to claim 1,wherein the cancer is selected from the group consisting of leukemia,prostate cancer, breast cancer and ovarian cancer.
 6. A method accordingto claim 1, wherein the binding agent specifically binds to anextracellular domain of OB-cadherin.
 7. A method according to claim 1,wherein the biological sample is selected from the group consisting ofblood, serum, urine, tumor biopsies, peritoneal fluid, cerebrospinalfluid, prostate secretions and fractions of the foregoing samples.
 8. Amethod for monitoring the progression of a cancer in a patient,comprising the steps of: (a) contacting a biological sample obtainedfrom a cancer patient at a first point in time with a binding agent thatspecifically binds to OB-cadherin; (b) detecting in the sample an amountof polypeptide that binds to the binding agent; (c) repeating steps (a)and (b) using a biological sample obtained from the patient at asubsequent point in time; and (d) comparing the amount of polypeptidedetected in step (c) to the amount detected in step (b) and therefrommonitoring the progression of the cancer in the patient.
 9. A methodaccording to claim 8 wherein the binding agent is a monoclonal antibody.10. A method according to claim 8 wherein the binding agent is apolyclonal antibody.
 11. A method according to claim 8, wherein thebinding agent comprises an OB-cadherin CAR sequence, or an analogue ormimetic thereof.
 12. A method according to claim 8, wherein the canceris selected from the group consisting of leukemia, prostate cancer,breast cancer and ovarian cancer.
 13. A method according to claim 8,wherein the binding agent specifically binds to an extracellular domainof OB-cadherin.
 14. A method according to claim 8, wherein thebiological sample is selected from the group consisting of blood, serum,urine, tumor biopsies, peritoneal fluid, prostate secretions andfractions of the foregoing samples.
 15. A method for evaluating themetastatic potential of a cancer in a patient, comprising the steps of:(a) contacting a biological sample obtained from a cancer patientafflicted with cancer with a binding agent that specifically binds toOB-cadherin; and (b) detecting in the sample an amount of polypeptidethat binds to the binding agent, relative to a predetermined cut-offvalue, and therefrom evaluating the metastatic potential of the cancerin the patient.
 16. A method according to claim 15 wherein the bindingagent is a monoclonal antibody.
 17. A method according to claim 15wherein the binding agent is a polyclonal antibody.
 18. A methodaccording to claim 15, wherein the binding agent comprises anOB-cadherin CAR sequence, or an analogue or mimetic thereof.
 19. Amethod according to claim 15, wherein the cancer is selected from thegroup consisting of leukemia, prostate cancer, breast cancer and ovariancancer.
 20. A method according to claim 15, wherein the binding agentspecifically binds to an extracellular domain of OB-cadherin.
 21. Amethod according to claim 15, wherein the biological sample is selectedfrom the group consisting of blood, serum, urine, tumor biopsies,peritoneal fluid, prostate secretions and fractions of the foregoingsamples.
 22. A diagnostic kit comprising: (a) one or more monoclonalantibodies that specifically bind to an OB-cadherin CAR sequence; and(b) a detection reagent.
 23. A kit according to claim 22 wherein themonoclonal antibodies are immobilized on a solid support.
 24. A kitaccording to claim 23 wherein the solid support comprisesnitrocellulose, latex or a plastic material.
 25. A kit according toclaim 22 wherein the detection reagent comprises a reporter group.
 26. Akit according to claim 25 wherein the reporter group is selected fromthe group consisting of radioisotopes, fluorescent groups, luminescentgroups, enzymes, biotin and dye particles.
 27. A method for determiningthe presence or absence of a metastatic cancer in a patient, comprisingthe steps of: (a) contacting a biological sample obtained from a patientwith an oligonucleotide that hybridizes to a polynucleotide that encodesOB-cadherin; and (b) detecting in the sample an amount of apolynucleotide that hybridizes to the oligonucleotide, relative to apredetermined cut-off value, and therefrom determining the presence orabsence of a metastatic cancer in the patient.
 28. A method according toclaim 27, wherein the amount of polynucleotide that hybridizes to theoligonucleotide is determined using polymerase chain reaction.
 29. Amethod according to claim 27, wherein the amount of polynucleotide thathybridizes to the oligonucleotide is determined using a hybridizationassay.
 30. A method according to claim 27, wherein the cancer isselected from the group consisting of leukemia, prostate cancer, breastcancer and ovarian cancer.
 31. A method according to claim 27, whereinthe biological sample is an RNA or cDNA preparation.
 32. A method formonitoring progression of a cancer in a patient, comprising the stepsof: (a) contacting a biological sample obtained from a cancer patientwith an oligonucleotide that hybridizes to a polynucleotide that encodesOB-cadherin; (b) detecting in the sample an amount of a polynucleotidethat hybridizes to the oligonucleotide; (c) repeating steps (a) and (b)using a biological sample obtained from the patient at a subsequentpoint in time; and (d) comparing the amount of polynucleotide detectedin step (c) with the amount detected in step (b) and therefrommonitoring progression of a cancer in the patient.
 33. A methodaccording to claim 32, wherein the amount of polynucleotide thathybridizes to the oligonucleotide is determined using polymerase chainreaction.
 34. A method according to claim 32, wherein the amount ofpolynucleotide that hybridizes to the oligonucleotide is determinedusing a hybridization assay.
 35. A method according to claim 32, whereinthe cancer is selected from the group consisting of leukemia, prostatecancer, breast cancer and ovarian cancer.
 36. A method according toclaim 32, wherein the biological sample is an RNA or cDNA preparation.37. A method for evaluating the metastatic potential of a cancer in apatient, comprising the steps of: (a) contacting a biological sampleobtained from a cancer patient with an oligonucleotide that hybridizesto a polynucleotide that encodes OB-cadherin; and (b) detecting in thesample an amount of a polynucleotide that hybridizes to theoligonucleotide, relative to a predetermined cut-off value, andtherefrom evaluating the metastatic potential of the cancer in thepatient.
 38. A method according to claim 37, wherein the amount ofpolynucleotide that hybridizes to the oligonucleotide is determinedusing polymerase chain reaction.
 39. A method according to claim 37,wherein the amount of polynucleotide that hybridizes to theoligonucleotide is determined using a hybridization assay.
 40. A methodaccording to claim 37, wherein the cancer is selected from the groupconsisting of leukemia, prostate cancer, breast cancer and ovariancancer.
 41. A method according to claim 37, wherein the biologicalsample is an RNA or cDNA preparation.
 42. A diagnostic kit, comprising:(a) an oligonucleotide that hybridizes to a polynucleotide that encodesOB-cadherin, or to a complement of such a polynucleotide; and (b) adiagnostic reagent for use in a polymerase chain reaction orhybridization assay.
 43. A diagnostic kit, comprising: (a) anoligonucleotide that hybridizes to a polynucleotide that encodesOB-cadherin, or to a complement of such a polynucleotide; and (b) asecond oligonucleotide 10-40 nucleotides in length. 44.-76. (Canceled)